Vous êtes sur la page 1sur 809

INDIAN

PHARMACOPOEIA
2007
Volume 3

THE INDIAN PHARMACOPOEIA COMMISSION


GHAZIABAD
INDIAN PHARMACOPOEIA 2007

Volume 3
CONTENTS

Monographs on Drug Substances, Dosage Forms and Pharmaceutical Aids


Monographs to Z ....

Monographs on Vaccines and Immunosera for Human Use ....


Monographs on Herbs and Herbal Products ....
Monographs on Blood and Blood-related Products ....
Monographs on Biotechnology Products ....
Monographs on Veterinary Products ....

Index ....
INDIAN PHARMACOPOEIA 2007 GENERAL NOTICES

GENERAL NOTICES

General Statements ....


Name ....
Official and Official Articles ....
Official Standards ....
Added Substances ....
Alternative Methods ....
Meanings of Terms ....
Provisions Applicable to Monographs and Test Methods ....
Expression of Contents ....
Expression of Concentrations ....
Abbreviated Statements ....
Weights and Measures ....
Monographs ....
General Monographs ....
Production ....
Manufacture of Drug Products ....
Excipients ....
Individual Monographs ....
Titles ....
Chemical Formulae ....
Atomic and Molecular Weights ....
Definitions ....
Statement of Contents ....
Descriptions ....
Identification ....
Tests and Assay ....
Tests ....
Other tests ....
Limits ....
Quantities ....

793
GENERAL NOTICES INDIAN PHARMACOPOEIA 2007

Apparatus ....
Reagents and Solutions ....
Indicators ....
Reference Substances ....
Tests Animals ....
Calculation of Results ....
Storage ....
Storage Containers ....
Labelling ....

794
IP 2007 GENERAL NOTICES

General Notices use but not necessarily to articles that may be sold under the
same name for other purposes.
The active pharmaceutical ingredients (drug substances),
General Statements excipients (pharmaceutical aids), pharmaceutical preparations
The General Notices provide the basic guidelines for the (dosage forms) and other articles described in the monographs
interpretation and application of the standards, tests, assays, are intended for human and veterinary use (unless explicitly
and other specifications of the Indian Pharmacopoeia (IP), as restricted to one of these uses).
well as to the statements made in the monographs and other The requirements given in the monographs are not framed to
texts of the Pharmacopoeia. provide against all possible impurities, contaminants or
A monograph is to be constructed in accordance with any adulterants; they provide appropriate limitation of potential
general monograph or notice or any appendix, note or other impurities only.
explanatory material that is contained in this Pharmacopoeia A preparation must comply throughout the shelf-life assigned
and that is applicable to that monograph. All statements to it by the manufacturer; for opened or broached containers
contained in the monograph, except where a specific general the maximum period of validity for use may sometimes be
notice indicates otherwise and with the exceptions given stated in the individual monograph. Nevertheless, the
hereafter, constitute standards for the official articles. An article responsibility for assigning the period of validity shall be
is not of pharmacopoeial quality unless it complies with all of with the manufacturer.
the requirements stated. Added Substances. An official substance, as distinguished
Exceptions to the General Notices do exist, and where they from an official preparation, contains no added substances
do, the wording in the individual monograph or an appendix except when specifically permitted in the individual monograph.
takes precedence and specifically indicates directions or the Unless otherwise specified in the individual monograph, or
intent. Thus, the specific wording of standards, tests, assays elsewhere in the General Notices, suitable substances may be
and other specifications is binding wherever deviations from added to an official preparation to enhance its stability,
the General Notices exist. Likewise, where there is no specific usefulness or elegance, or to facilitate its preparation. Such
mention to the contrary, the General Notices apply. auxiliary substances shall be harmless in the amounts used,
shall not exceed the minimum quantity required to provide
Name. The full name or title of this book, including addenda
their intended effect, shall not impair the therapeutic efficacy
thereto, is Indian Pharmacopoeia 2007, abbreviated to IP 2007.
or the bioavailability or safety of the preparation and shall not
In the texts, the term “Pharmacopoeia” or “IP” without
interfere with the tests and assays prescribed for determining
qualification means the Indian Pharmacopoeia 2007 and any
compliance with the official standards. Particular care should
addenda thereto.
be taken to ensure that such substances are free from harmful
Official and Official Articles. The word ‘official’ wherever organisms. The freedom to the manufacturers to add auxiliary
used in this Pharmacopoeia or with reference thereto, is substances imposes on them the responsibility of satisfying
synonymous with ‘pharmacopoeial’, with ‘IP’ and with the licensing authorities on the purpose of the addition and
‘compendial’. The designation IP in conjunction with the the innocuity of such substances.
official title on the label of an article is an indication that the Alternative Methods. The tests and assays described are the
article purports to comply with IP standards. official methods upon which the standards of the
The following terms are used where the articles for which Pharmacopoeia are based. Alternative methods of analysis
monographs are provided are to be distinguished. may be used for control purposes, provided that the methods
used are shown to give results of equivalent accuracy and
An official substance is a single drug or a drug entity or a enable an unequivocal decision to be made as to whether
pharmaceutical aid for which the monograph title includes no compliance with the standards of the monographs would be
indication of the nature of a dosage form. achieved if the official methods were used. Automated
An official preparation is a drug product (dosage form) and is procedures utilising the same basic chemistry as the test
the finished or partially finished preparation or product of one procedures given in the monograph may also be used to
or more official substances formulated for use on the patient. determine compliance. Such alternative or automated
procedures must be validated.
An article is an item for which a monograph is provided,
whether an official substance or an official preparation. In the event of doubt or dispute, the methods of analysis of
the Pharmacopoeia are alone authoritative and only the result
Official Standards. The requirements stated in the obtained by the procedure given in this Pharmacopoeia is
monographs apply to articles that are intended for medicinal conclusive.

795
GENERAL NOTICES IP 2007

Meanings of Terms — per cent v/v (percentage, volume in volume) expressing


Alcohol. The term “alcohol” without qualification means the number of millilitres of substance in 100 millilitres of
ethanol (95 per cent). Other dilutions of ethanol are indicated final product.
by the term “alcohol” or “alcohol” followed by a statement of The expression “parts per million” refers to the weight in
the percentage by volume of ethanol (C2H6O) required. weight, unless otherwise stated.
Desiccator. A tightly-closed container of suitable size and Where the content of a substance is expressed in terms of the
design that maintains an atmosphere of low moisture content chemical formula for that substance an upper limit exceeding
by means of silica gel or phosphorus pentoxide or other 100 per cent may be stated. Such an upper limit applies to the
suitable desiccant. result of the assay calculated in terms of the equivalent content
of the specified chemical formula. For example, the statement
Drying and ignition to constant weight. Two consecutive
‘contains not less than 99.0 per cent and not more than 101.0
weighings after the drying or igniting operations do not differ
per cent of C7H6O2 implies that the result of the assay is not
by more than 0.5 mg, the second weighing following an
less than 99.0 per cent and not more than 101.0 per cent,
additional period of drying or of ignition respectively
calculated in terms of the equivalent content of C7H6O2.
appropriate to the nature and quantity of the residue.
Where the result of an assay or test is required to be calculated
Ethanol. The term “ethanol” without qualification means
with reference to the dried, anhydrous, ignited substance, or
anhydrous ethanol or absolute alcohol.
the substance free from solvent, the determination of loss on
Filtration. Unless otherwise stated, filtration is the passing of drying, water content, loss on ignition, content of the specified
a liquid through a suitable filter paper or equivalent device solvent, respectively is carried out by the method prescribed
until the filtrate is clear. in the relevant test in the monograph.
Freshly prepared. Made not more than 24 hours before it is Expression of Concentrations. The following expressions in
issued for use. addition to the ones given under Expression of Content are
also used:
Label. Any printed packing material, including package inserts
that provide information on the article. — per cent w/v (percentage, weight in volume) expressing
the number of grams of substance in 100 millilitres of
Negligible. A quantity not exceeding 0.50 mg. product
Solution. Where the name of the solvent is not stated, — per cent v/w (percentage, volume in weight) expressing
“solution” implies a solution in water. The water used complies the number of millilitres of substance in 100 grams of
with the requirements of the monograph on Purified Water. product.
The term ‘distilled water’ indicates Purified Water prepared by
distillation. Usually, the strength of solutions of solids in liquids is
expressed as percentage weight in volume, of liquids in liquids
Temperature. The symbol º used without qualification as percentage volume in volume, of solids in semi-solid bases
indicates the use of the Celsius thermometric scale. (e.g. creams) and of gases in liquids as percentage weight in
Water. If the term is used without qualification it means Purified weight.
Water of the Pharmacopoeia. The term ‘distilled water’ When the concentration of a solution is expressed as parts of
indicates Purified Water prepared by distillation. dissolved substance in parts of solution, it means parts by
weight (g) of a solid in parts by volume (ml) of the final solution;
Water-bath. A bath of boiling water unless water at another
as parts by weight (g) of a gas in parts by weight (g) of the
temperature is indicated. Other methods of heating may be
final solution.
used provided the required temperature is approximately
maintained but not exceeded. When the concentration of a solution is expressed in molarity
designated by the symbol M preceded by a number, it denotes
Provisions Applicable To Monographs and Test Methods the number of moles of the stated solute contained in sufficient
Purified Water (unless otherwise stated) to produce 1 litre of
Expression of Content. Where the content of a substance is solution.
defined, the expression “per cent” is used according to
circumstances with one of two meanings: Abbreviated Statements. Incomplete sentences are employed
in parts of the monographs for directness and brevity (for
— per cent w/w (percentage, weight in weight) expressing example, Iodine Value. Not more than ……; Relative Density.
the number of grams of substance in 100 grams of final …….to……..) Where the tests are abbreviated, it is to be
product, understood that the test method referred to in brackets

796
IP 2007 GENERAL NOTICES

provides the method to be followed and that the values Excipients. Any substance added in preparing an official
specified are the applicable limits. preparation shall be innocuous, shall have no adverse influence
in the therapeutic efficacy of the active ingredients and shall
Weights and Measures. The metric system of weights and
not interfere with the tests and assays of the Pharmacopoeia.
measures is employed in the Pharmacopoeia. All measures are
Care should be taken to ensure that such substances are free
required to be graduated at 25º and all measurements in tests
from harmful organisms.
and assays, unless otherwise stated, are to be made at that
temperature. Graduated glass apparatus used in analytical Individual Monographs
operations shall comply with the requirements stated in
Chapter 2.1.6 Drug products that are the subject of an individual monograph
are also required to comply with the tests given in the general
monographs.
Monographs
Titles. The main title for a drug substance is the International
General Monographs Non-proprietary Name (INN) approved by the World Health
Organization. Subsidiary names and synonyms have also been
General monographs on dosage forms include requirements given in some cases; where included, they have the same
of general application and apply to all preparations within the significance as the main title.
scope of the Introduction section of the general monograph,
except where a preamble limits the application. The The main titles of drug products are the ones commonly
requirements are not necessarily comprehensive for a given recognised in practice. Synonyms drawn from the full non-
specific preparation; additional requirements may sometimes proprietary name of the active ingredient or ingredients have
be given in the individual monograph for it. also been given. Where, however, a product contains one or
the other of different salts of an active molecule, the main title
Production. Statements given under the heading Production is based on the full name of the active ingredient. For example,
relate to particular aspects of the manufacturing process and Chloroquine Phosphate Tablets and Chloroquine
are not necessarily comprehensive. However, they are SulphateTablets.
mandatory instructions to manufacturers. They may relate,
for example, to source materials, to the manufacturing process Chemical Formulae. When the chemical structure of an official
and its validation and control, to any in-process testing that substance is known or generally accepted, the graphic and
is to be carried out by the manufacturer on the final product molecular formulae are normally given at the beginning of the
either on selected batches or on each batch prior to release. monograph for information. This information refers to the
All this cannot be verified on a sample of the final product by chemically pure substance and is not to be regarded as an
an independent analyst. It is for the licensing authority to indication of the purity of the official material. Elsewhere, in
verify that the instructions have been followed. statement of purity and strength and in descriptions of
processes of assay, it will be evident from the context that the
The absence of a section on Production does not imply that formulae denote the chemically pure substances.
attention to features such as those given above is not required.
An article described in a monograph of the Pharmacopoeia is Where the absolute stereochemical configuration is specified,
to be manufactured in accordance with the principles of good the International Union of Pure and Applied Chemistry
manufacturing practice and in accordance with the (IUPAC) R/S and E/Z systems of designation have been used.
requirements of the Drugs and Cosmetics Rules, 1945. The If the substance is an enantiomer of unknown absolute
general principles applicable to the manufacture and quality stereochemistry, the sign of the optical rotation, as determined
assurance of drugs and preparations meant for human use in the solvent and under the conditions specified in the
apply equally to veterinary products as well. monograph, has been attached to the systematic name. An
indication of sign of rotation has also been given where this is
Manufacture of Drug Products. The opening definitive incorporated in a trivial name that appears on an IUPAC
statement in certain monographs for drug products is given in preferred list.
terms of the active ingredient(s) only. Any ingredient(s) other
than those included in the statement, must comply with the Atomic and Molecular Weights. The atomic weight or
general notice on Excipients and the product must conform to molecular weight is shown , as and when appropriate at the
the Pharmacopoeial requirements. top right hand corner of the monograph. The atomic and
molecular weights and graphic formulae do not constitute
Official preparations are prepared only from ingredients that analytical standards for the substances described.
comply with the requirements of the pharmacopoeial
monographs for those individual ingredients for which Definition. The opening statement of a monograph is one
monographs are provided. that constitutes an official definition of the substance,

797
GENERAL NOTICES IP 2007

preparation or other article that is the subject of the are not framed to take into account all possible impurities. It is
monograph. In certain monographs for pharmaceutical not to be presumed, for example, that an impurity that is not
preparations the statement is given in terms of the principal detectable by means of the prescribed tests is tolerated.
ingredient(s). Material found to contain such an impurity is not of
In monographs on vegetable drugs, the definition indicates pharmacopoeial quality if the nature or amount of the impurity
whether the subject of the monograph is, for example, the found is incompatible with good pharmaceutical practice.
whole drug or the drug in powdered form. Pharmacopoeial methods and limits should be used merely as
Certain pharmaceutical substances and other articles are compliance requirements and not as requirements to guarantee
defined by reference to a particular method of manufacture. A total quality assurance. Tests and assays are prescribed for
statement that a substance or article is prepared or obtained the minimum sample available on which the attributes of the
by a certain method constitutes part of the official definition article should be measured. Assurance of quality must be
and implies that other methods are not permitted. A statement ensured by the manufacturer by the use of statistically valid
that a substance may be prepared or obtained by a certain sampling and testing programmes.
method, however, indicates that this is one possible method Tests. Unless otherwise stated, the assays and tests are carried
and does not imply that other methods are not permissible. out at a temperature between 20º and 30º.
Statement of content. The limits of content stated are those Where it is directed that an analytical operation is to be carried
determined by the method described under Assay. out ‘in subdued light’, precautions should be taken to avoid
Description. The statements under the heading Description exposure to direct sunlight or other strong light. Where a
are not to be interpreted in a strict sense and are not to be procedure is directed to be performed ‘protected from light’
regarded as official requirements. precautions should be taken to exclude actinic light by the
Solubility. Statements on solubility are given in Chapter 2.4.26 use of low-actinic glassware, working in a dark room or similar
and are intended as information on the approximate solubility procedures.
at a temperature between 15º and 30º, unless otherwise stated, For preparations other than those of fixed strength, the
and are not to be considered as official requirements. However, quantity to be taken for a test or an assay is usually expressed
a test for solubility stated in a monograph constitutes part of in terms of the active ingredient. This means that the quantity
the standards for the substance that is the subject of that of the active ingredient expected to be present and the quantity
monograph. of the preparation to be taken are calculated from the strength
stated on the label.
Test Methods
Other Tests. In the monographs on dosage forms and certain
References to general methods of testing are indicated by test preparations, under the sub-heading ‘Other tests’ it is stated
method numbers in brackets immediately after the heading of that the article complies with the tests stated under the general
the test or at the end of the text. monograph of the relevant dosage form or preparation. Details
Identification. The tests given under the heading Identification of such tests are provided in the general monographs.
are not necessarily sufficient to establish absolute proof of Limits. The limits given are based on data obtained in normal
identity. They provide a means of verifying that the identity analytical practice. They take into account normal analytical
of the material under examination is in accordance with the errors, of acceptable variations in manufacture and of
label on the container. deterioration to an extent that is acceptable. No further
In certain monographs alternative series of identification tests tolerances are to be applied to the limits for determining whether
are given; compliance with either one or the other set of tests or not the article under examination complies with the
is adequate to verify the identity of the article. requirements of the monograph.
When tests for infrared absorption are applied to material Quantities. Unless otherwise stated, the quantities to be taken
extracted from formulated preparations, strict concordance for assays, limit tests and other tests are of the substance
with the specified reference spectrum may not always be under examination.
possible, but nevertheless a close resemblance between the In tests with numerical limits and assays, the quantity stated
spectrum of the extracted material and the specified reference to be taken for testing is approximate. The amount actually
spectrum should be achieved. used, which may deviate by not more than 10 per cent from
that stated, is accurately weighed or measured and the result
Tests and Assays
of analysis is calculated from this exact quantity. In tests where
The tests and assays are the official methods upon which the the limit is not numerical but usually depends upon
standards of the Pharmacopoeia depend. The requirements comparison with the behaviour of a reference in the same

798
IP 2007 GENERAL NOTICES

conditions, the stated quantity is taken for testing. Reagents Indian Pharmacopoeia Commission (IPC). They are the official
are used in the prescribed amounts. standards to be used in cases of arbitration. Secondary
Quantities are weighed or measured with an accuracy Standards (Working Standards) may be used for routine
commensurate with the indicated degree of precision. For analysis, provided they are standardized at regular intervals
weighings, the precision is plus or minus 5 units after the last against the Reference Substances
figure stated. For example, 0.25 g is to be interpreted as 0.245 Biological Reference Substances, also abbreviated to IPRS
g to 0.255 g. For the measurement of volumes, if the figure and Standard Preparations of antibiotics are issued by
after the decimal point is a zero or ends in a zero, e.g. 10.0 ml 0r agencies authorised by the IPC. They are standardized against
0.50 ml, the volume is measured using a pipette, a volumetric the International Standards and Reference Preparations
flask or a burette, as appropriate; in other cases, a graduated established by the World Health Organization (WHO). The
measuring cylinder or a graduated pipette may be used. potency of these preparations is expressed in International
Volumes stated in microlitres are measured using a micropipette Units.
or microsyringe.
Reference spectra are published by the IPC and they are
The term ‘transfer’ is used generally to indicate a quantitative accompanied by information concerning the conditions used
operation. for sample preparation and recording of the spectra.
Apparatus. Measuring and weighing devices and other Test animals. Unless otherwise directed, animals used in a
apparatus are described in the chapter entitled ‘Apparatus for test or an assay shall be healthy and are drawn from a uniform
Tests and Assays’. A specification for a definite size or type stock, and have not previously been treated with any material
of container or apparatus in a test or assay is given merely as that will interfere with the test or the assay.
a recommendation. Calculation of results. In determining compliance with a
Unless otherwise stated, comparative tests are carried out numerical limit in assay or test, the result should be calculated
using identical tubes of colourless, transparent, neutral glass to one decimal place more than the significant figures stated
with a flat base, commonly known as Nessler cylinders. and then rounded up or down as follows: if the last figure
calculated is 5 to 9, the preceding figure is increased by 1; if it
Reagents and Solutions. The reagents required for the tests
is 4 or less, the preceding figure is left unchanged.
and assays of the Pharmacopoeia are defined in the various
chapters showing their nature, degree of purity and the Storage. Statements under the side-heading Storage constitute
strengths of the solutions to be made from them. The non-mandatory advice. The articles of the Pharmacopoeia are
requirements set out are not intended to imply that the materials to be stored under conditions that prevent contamination and,
are suitable for use in medicine; regents not covered by as far as possible, deterioration. Precautions that should be
monographs in the pharmacopoeia shall not be claimed to be taken in relation to the effects of the atmosphere, moisture,
of IP quality. heat and light are indicated, where appropriate, in the individual
monograph.
The term ‘analytical reagent grade of commerce’ implies that
the chemical is of a high degree of purity wherein the limits of Specific directions are given in some monographs with respect
various impurities are known. Where it is directed to use a to the temperatures at which Pharmacopoeial articles should
‘general laboratory reagent grade of commerce’ it is intended be stored, where it is considered that usage at a lower or
that a chemically pure grade material, not necessarily required higher temperature may produce undesirable results. The
to be tested for limiting or absence of certain impurities, is to storage conditions are defined by the following terms:
be used. — Store in a dry, well-ventilated place at a temperature not
Indicators. Where the use of an indicator solution is mentioned exceeding 30º
in an assay or test, approximately 0.1 ml of the solution shall — Store in a refrigerator (2º to 8º). Do not freeze
be added, unless otherwise directed.
— Store in a freezer (-2º to -18º)
Reference Substances. Certain monographs require the use — Store in a deep freezer (Below -18º)
of a chemical reference substance or a biological reference
preparation or a reference spectrum These are authentic Storage conditions not related to temperature are indicated in
specimens chosen and verified on the basis of their suitability the following terms:
for intended use as prescribed in the Pharmacopoeia and are — Store protected from light
not necessarily suitable in other circumstances. — Store protected from light and moisture
IP Reference Substances, abbreviated to IPRS (and referred Where no specific storage directions or limitations are given
to as RS in the individual monographs) are issued by the in the monograph or by the manufacturer, it is to be understood

799
GENERAL NOTICES IP 2007

that the storage conditions include protection from moisture, of being tightly closed, and re-closed after use.
freezing and excessive heat (any temperature above 40º).
In certain cases, special requirements of pack have been
Storage Containers. The requirements, guidance and indicated in some monographs under Storage, using
information on containers for pharmaceutical use are given in expressions that have been defined in chapter 6.1.
the chapter entitled Containers (6.1) Labelling. The labelling of drugs and pharmaceuticals is
In general, an article should be packed in a well-closed governed by the Drugs and Cosmetics Rules, 1945. The
container i.e. one that protects the contents from statements that are given in the monographs under the side-
contamination by extraneous solids, liquids or vapours and heading ‘Labelling’ are not comprehensive. Only those that
from loss of the article under normal conditions of handling are necessary to demonstrate compliance or otherwise with
and storage. the monograph have been given and they are mandatory. For
example, in the monograph on Betamethasone Sodium Tablets
Where, additionally, loss or deterioration of the article from
the labelling statement is “The label states the strength in
effervescence, deliquescence or evaporation under normal
terms of the equivalent amount of betamethasone”. Any other
conditions of storage is likely, the container must be capable
statements are included as recommendations.

800
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

DRUG SUBSTANCES, DOSAGE FORMS


AND
PHARMACEUTICAL AIDS

N to Z ..................................................................................................................

801
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

N
Nalidixic Acid ....
Nalidixic Acid Tablets ....
Nalorphine Hydrochloride ....
Nalorphine Injection ....
Nandrolone Decanoate ....
Nandrolone Decanoate Injection ....
Nandrolone Phenylpropionate ....
Nandrolone Phenylpropionate Injection ....
Naphazoline Nitrate ....
Nelfinavir Mesylate ....
Nelfinavir Mesylate Oral Powder ....
Nelfinavir Tablets ....
Neomycin Sulphate ....
Neomycin Eye Drops ....
Neomycin Eye Ointment ....
Neostigmine Bromide ....
Neostigmine Tablets ....
Neostigmine Methylsulphate ....
Neostigmine Injection ....
Nevirapine ....
Nevirapine Oral Suspension ....
Nevirapine Tablets ....
Niclosamide ....
Niclosamide Tablets ....
Nicotinamide ....
Nicotinamide Tablets ....
Nicotinic Acid ....
Nicotinic Tablets ....
Nicoumalone ....
Nicoumalone Tablets ....

803
MONOGRAPHS INDIAN PHARMACOPOEIA 2007 2007

Nifedipine ....
Nifedipine Capsules ....
Nifedipine Sustained release-Tablets ....
Nifedipine Tablets ....
Nikethamide ....
Nikethamide Injection ....
Nitrazepam ....
Nitrazepam Tablets ....
Nitrofurantoin ....
Nitrofurantoin Tablets ....
Nitrofurazone ....
Nitrous Oxide ....
Noradrenaline Bitartrate ....
Noradrenaline Bitartrate Injection ....
Norethisterone ....
Norethisterone Tablets ....
Norfloxacin ....
Norfloxacin Eye Drops ....
Norfloxacin Tablets ....
Norgestrel ....
Norgestrel And Ethinyloestradiol Tablets ....
Nortriptyline Hydrochloride ....
Nortriptyline Tablets ....
Noscapine ....
Noscapine Linctus ....
Novobiocin Sodium ....
Nystatin ....
Nystatin Ointment ....
Nystatin Pessaries ....
Nystatin Tablets ....

804
IP 2007 NALIDIXIC ACID TABLETS

Nalidixic Acid Reference solution (b). A 0.0008 per cent w/v solution of the
substance under examination in dichloromethane.
CH3 Reference solution (c). A 0.1 per cent w/v solution of nalidixic
acid RS in dichloromethane.
H3C N N Apply to the plate 10 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
COOH Any secondary spot in the chromatogram obtained with test
O solution (a) is not more intense than the spot in the
chromatogram obtained with reference solution (a) and not
more than one such spot is more intense than the spot in the
C12H12N2O3 Mol. Wt. 232.2
chromatogram obtained with reference solution (b).
Nalidixic Acid is 1-ethyl-1,4-dihydro-7-methyl-4-oxo-1,8-
naphthyridine-3-carboxylic acid. Heavy metals (2.3.13). 1.0 g complies with the limit test for
heavy metals, Method B (20 ppm).
Nalidixic Acid contains not less than 99.0 per cent and not
more than 101.0 per cent of C12H12N2O3, calculated on the Sulphated ash (2.3.18) Not more than 0.1 per cent.
dried basis. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Description. A white to slightly yellow, crystalline powder. on 1.0 g by drying in an oven at 105°.
Assay. Weigh accurately about 0.15 g, dissolve in 10 ml of
Identification dichloromethane, add 30 ml of 2-propanol and 10 ml of carbon
dioxide-free water and titrate with 0.1 M ethanolic sodium
Test A may be omitted if tests B, C and D are carried out. Tests
hydroxide, determining the end-point potentiometrically
B, C and D may be omitted if test A is carried out.
(2.4.25) and using a glass electrode as the indicator electrode
A. Determine by infrared absorption spectrophotometry (2.4.6). and a silver-silver chloride reference electrode with a sleeve
Compare the spectrum with that obtained with nalidixic acid diaphragm or a capillary tip filled with a saturated solution of
RS or with the reference spectrum of nalidixic acid. lithium chloride in ethanol. Throughout the titration keep
B. When examined in the range 230 nm to 360 nm (2.4.7), a the temperature of the solution at 15° to 20° and pass a current
0.0005 per cent w/v solution in 0.1 M sodium hydroxide shows of nitrogen through the solution.
absorption maxima at about 258 nm and 334 nm; ratio of the 1 ml of 0.1 M ethanolic sodium hydroxide is equivalent to
absorbance at about 258 nm to that at about 334 nm, 2.2 to 2.4. 0.02322 g of C12H12N2O3.
C. In the test for Related substances, the principal spot in the Storage. Store protected from light and moisture.
chromatogram obtained with test solution (b) corresponds to
that in the chromatogram obtained with reference solution (c).
D. Dissolve 0.1 g in 2 ml of hydrochloric acid and add 0.5 ml
of a 10 per cent w/v solution of 2-naphthol in ethanol (95 per Nalidixic Acid Tablets
cent); an orange-red colour develops.
Nalidixic Acid Tablets contain not less than 95.0 per cent and
Tests not more than 105.0 per cent of the stated amount of nalidixic
acid, C12H12N2O3.
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel HF254. Identification
Mobile phase. A mixture of 70 volumes of ethanol (95 per To a quantity of the powdered tablets containing 1 g of Nalidixic
cent), 20 volumes of dichloromethane and 10 volumes of 5 M Acid add 50 ml of chloroform, shake for 15 minutes, filter and
ammonia. evaporate the filtrate to dryness. The residue, after drying at
Test solution (a). Dissolve 0.2 g of the substance under 105°, complies with the following tests.
examination in 10 ml of dichloromethane. A. Determine by infrared absorption spectrophotometry (2.4.6).
Test solution (b). A 0.1 per cent w/v solution of the substance Compare the spectrum with that obtained with nalidixic acid
under examination in dichloromethane. RS or with the reference spectrum of nalidixic acid.
Reference solution (a). A 0.002 per cent w/v solution of the B. When examined in the range 230 nm to 360 nm (2.4.7), a
substance under examination in dichloromethane. 0.0005 per cent w/v solution in 0.1 M sodium hydroxide shows

805
NALORPHINE HYDROCHLORIDE IP 2007

absorption maxima at about 258 nm and 334 nm; ratio of the Nalorphine Hydrochloride contains not less than 97.0 per cent
absorbance at about 258 nm to that at about 334 nm, 2.2 to 2.4. and not more than 103.0 per cent of C19H21NO3,HCl, calculated
on the dried basis.
Tests
Description. A white or almost white, crystalline powder;
Related substances. Determine by thin-layer chromatography odourless. It slowly darkens on exposure to air and light.
(2.4.17), coating the plate with silica gel HF254.
Identification
Mobile phase. A mixture of 70 volumes of ethanol (95 per
cent), 20 volumes of dichloromethane and 10 volumes of Test A may be omitted if tests B, C, D and E are carried out.
5 M ammonia. Tests C and D may be omitted if tests A, B and E are carried
out.
Test solution. Shake a quantity of the powdered tablets
containing 0.1 g of Nalidixic Acid with 50 ml of chloroform for A. Determine by infrared absorption spectrophotometry (2.4.6).
15 minutes, filter, evaporate the filtrate to dryness and dissolve Compare the spectrum with that obtained with nalorphine
the residue in 5 ml of chloroform. hydrochloride RS.
Reference solution. Dilute 1 volume of the test solution to B. When examined in the range 230 nm to 360 nm (2.4.7), a
200 volumes with chloroform. 0.01 per cent w/v solution in 0.1 M sodium hydroxide shows
Apply to the plate 10 µl of each solution. After development, an absorption maximum only at about 298 nm; absorbance at
dry the plate in air and examine in ultraviolet light at 254 nm. about 298 nm, about 0.6.
Any secondary spot in the chromatogram obtained with the C. To 10 ml of a 2 per cent w/v solution add 0.05 ml of dilute
test solution is not more intense than the spot in the ammonia solution; a white precipitate soluble in sodium
chromatogram obtained with the reference solution. hydroxide solution is produced.
Other Tests. Complies with the tests stated under Tablets. D. Dissolve 2 mg in 2 ml of water, add 0.15 ml of potassium
Assay. Weigh and powder 20 tablets. Weigh accurately a ferricyanide solution containing, in each ml, 0.05 ml of ferric
quantity of the powder containing about 0.1 g of Nalidixic chloride solution; a deep bluish green colour is produced
Acid, add 150 ml of 0.1 M sodium hydroxide, shake for 3 immediately.
minutes, dilute to 200.0 ml with 0.1 M sodium hydroxide, mix E. Gives reaction A of chlorides (2.3.1).
and allow to stand for 15 minutes. Dilute 2.0 ml of the solution
to 100.0 ml with water and measure the absorbance of the Tests
resulting solution at the maximum at about 334 nm (2.4.7),
using 0.1 M sodium hydroxide as the blank. Calculate the Melting range (2.4.21). 260° to 263°.
content of C12H12N2O3 taking 494 as the specific absorbance Acidity. Dissolve 0.2 g in 10 ml of freshly boiled and cooled
at 334 nm. water and titrate with 0.02 M sodium hydroxide using methyl
Storage. Store protected from light and moisture. red solution as indicator; not more than 0.2 ml of 0.02 M
sodium hydroxide is required to change the colour of the
solution.
Specific optical rotation (2.4.22). –122° to –125°, determined
Nalorphine Hydrochloride in a 2.0 per cent w/v solution.
Sulphated ash (2.3.18). Not more than 0.1 per cent.
HO
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
on 1.0 g by drying in an oven at 100° at a pressure not exceeding
O 0.7 kPa for 2 hours.
N , HCl
Assay. Weigh accurately about 25 mg and dissolve in sufficient
H CH2
water to produce 250 ml. Measure the absorbance of the
HO resulting solution at the maximum at about 285 nm (2.4.7).
Calculate the content of C19H21NO3,HCl from the absorbance
C19H21NO3,HCl Mol. Wt. 347.8 obtained by repeating the operation with nalorphine
Nalorphine Hydrochloride is 17-allyl-7,8-didehydro-4,5α- hydrochloride RS in place of the substance under
epoxymorphinan-3,6α-diol hydrochloride. examination.

806
IP 2007 NANDROLONE DECANOATE

Storage. Store protected from light and moisture. Nandrolone Decanoate

O
Nalorphine Injection
H3C O CH2(CH2)7CH3
Nalorphine Hydrochloride Injection
Nalorphine Injection is a sterile solution of Nalorphine H H
Hydrochloride in Water for Injections containing suitable
H H
buffering agents.
O
Nalorphine Injection contains not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of nalorphine
hydrochloride, C19H21NO3,HCl. C28H44O3 Mol. Wt. 428.7
Nandrolone Decanoate is 3-oxo-4-estren-17β-yl decanoate.
Identification Nandrolone Decanoate contains not less than 97.0 per cent
A. To a volume containing 50 mg of Nalorphine Hydrochloride and not more than 103.0 per cent of C28H44O3, calculated on
add dilute ammonia solution until the solution is alkaline and the dried basis.
extract with 25 ml of a mixture of 1 volume of ethanol (95 per Description. A white to creamy-off white, crystalline powder;
cent) and 3 volumes of chloroform and evaporate the extract odour, faint and characteristic.
to dryness. Dry the residue at a pressure not exceeding 2 kPa.
The residue complies with the following test. Identification
Determine by infrared absorption spectrophotometry (2.4.6). A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with nalorphine Compare the spectrum with that obtained with nandrolone
hydrochloride RS. decanoate RS or with the reference spectrum of nandrolone
B. To a volume containing 0.1 g of Nalorphine Hydrochloride decanoate.
add 0.05 ml of dilute ammonia solution; a white precipitate B. When examined in the range 230 nm to 360 nm (2.4.7), a
soluble in sodium hydroxide solution is produced. 0.001 per cent w/v solution in ethanol (95 per cent) shows an
absorption maximum only at about 239 nm; absorbance at
C. Gives reaction A of chlorides (2.3.1).
about 239 nm, about 0.41.
Tests C. Dissolve 25 mg in 1 ml of methanol, add 2 ml of
semicarbazide acetate solution, heat under a reflux condenser
pH (2.4.24). 6.0 to 7.5. for 30 minutes and cool; the precipitate, after recrystallisation
Other Tests. Complies with the tests stated under Parenteral from ethanol (95 per cent), melts at about 175° (2.4.21).
Preparations (Injections).
Tests
Assay. Transfer an accurately measured volume containing
about 10 mg of Nalorphine Hydrochloride to a separating Specific optical rotation (2.4.22). +32.0 ° to +36.0°, determined
funnel, add 1 ml of dilute hydrochloric acid and dilute to in a 2.0 per cent w/v solution in dioxan.
10 ml with water. Extract with five successive quantities, each Related substances. Determine by thin-layer chromatography
of 5 ml, of chloroform, allowing the layers to separate before (2.4.17), coating the plate with silica gel GF254.
drawing off each chloroform extract and discard the chloroform
Mobile phase. A mixture of 70 volumes of heptane and
extracts. Transfer the aqueous layer to a 100-ml volumetric
30 volumes of acetone.
flask with the aid of small quantities of water and dilute to
volume with water. Measure the absorbance of the resulting Test solution. Dissolve 0.1 g of the substance under
solution at the maximum at about 285 nm (2.4.7). Calculate examination in 10 ml of chloroform.
the content of C19H21NO3,HCl from the absorbance obtained Reference solution (a). A 0.005 per cent w/v solution of the
by repeating the operation with nalorphine hydrochloride substance under examination in chloroform.
RS.
Reference solution (b). A 0.01 per cent w/v solution of
Storage. Store protected from light. nandrolone RS in chloroform.

807
NANDROLONE DECANOATE INJECTION IP 2007

Apply to the plate 5 µl of each solution. After development, Tests


dry the plate in air and examine in ultraviolet light at 254 nm. In
the chromatogram obtained with the test solution any spot Other Tests. Complies with the tests stated under Parenteral
corresponding to nandrolone is not more intense than the Preparations (Injections).
spot in the chromatogram obtained with reference solution Assay. To an accurately measured volume containing about
(b) and any other secondary spot is not more intense than the 0.1 g of Nandrolone Decanoate add sufficient chloroform to
spot in the chromatogram obtained with reference solution produce 100.0 ml. Dilute 3.0 ml of the solution to 50.0 ml with
(a). chloroform. To 5.0 ml of this solution add 10 ml of isoniazid
Sulphated ash (2.3.18). Not more than 0.1 per cent. solution and sufficient methanol to produce 20.0 ml. Allow to
stand for 45 minutes and measure the absorbance of the
Loss on drying (2.4.19). Not more than 0.5 per cent, determined resulting solution at the maximum at about 380 nm (2.4.7),
on 1.0 g by drying over phosphorus pentoxide at a pressure using as the blank 5 ml of chloroform treated in the same
not exceeding 0.7 kPa for 4 hours. manner. Calculate the content of C28H44O3 from the absorbance
Assay. Weigh accurately about 10 mg and dissolve in sufficient obtained by repeating the operation using a suitable quantity
ethanol (95 per cent) to produce 100.0 ml. Dilute 5.0 ml to of nandrolone RS.
50.0 ml with ethanol (95 per cent) and measure the absorbance 1 mg of C18H26O2 is equivalent to 1.562 mg of C28H44O3.
of the resulting solution at the maximum at about 239 nm (2.4.7).
Calculate the content of C28H44O3 taking 407 as the specific Storage. Store protected from light.
absorbance at 239 nm.
Storage. Store protected from light and moisture.
Nandrolone Phenylpropionate
Nandrolone Phenpropionate
Nandrolone Decanoate Injection O
Nandrolone Decanoate Injection is a sterile solution of
Nandrolone Decanoate in Ethyl Oleate or other suitable ester, H3C O
in a suitable fixed oil or in any mixture of these.
H H
Nandrolone Decanoate Injection contains not less than 90.0
per cent and not more than 110.0 per cent of the stated amount H H
of nandrolone decanoate, C28H44O3. O
Identification C27H34O3 Mol.Wt. 406.6
Determine by thin-layer chromatography (2.4.17), coating the Nandrolone Phenylpropionate is 3-oxo-4-estren-17β-yl 3-
plate with silica gel GF254. phenylpropionate.
Mobile phase. A mixture of 70 volumes of heptane and Nandrolone Phenylpropionate contains not less than 97.0 per
30 volumes of acetone. cent and not more than 103.0 per cent of C27H34O3, calculated
Test solution. Dilute a suitable volume of the injection with on the dried basis.
carbon tetrachloride to give a solution containing 0.5 per- Description. A white to creamy-white, crystalline powder;
cent w/v solution of Nandrolone Decanoate. odour, characteristic.
Reference solution. A 0.5 per cent w/v solution of nandrolone
Identification
decanoate RS in carbon tetrachloride.
Apply to the plate 5 µl of each solution. After development, A. Determine by infrared absorption spectrophotometry (2.4.6).
dry the plate in air until the odour of solvent is no longer Compare the spectrum with that obtained with nandrolone
detectable, spray with a 10 per cent v/v solution of sulphuric phenylpropionate RS or with the reference spectrum of
acid in ethanol (95 per cent), heat at 105° for 30 minutes and nandrolone phenylpropionate.
examine in ultraviolet light at 365 nm. The principal spot in the B. When examined in the range 230 nm to 360 nm (2.4.7), a
chromatogram obtained with the test solution corresponds to 0.001 per cent w/v solution in ethanol (95 per cent) shows an
that in the chromatogram obtained with the reference solution. absorption maximum only at about 240 nm; absorbance at
Ignore any subsidiary spots due to the vehicle. about 240 nm, about 0.43.

808
IP 2007 NANDROLONE PHENYLPROPIONATE INJECTION

C. Dissolve 25 mg in 1 ml of methanol, add 2 ml of Identification


semicarbazide acetate solution, heat under a reflux condenser
for 30 minutes and cool; the precipitate, after recrystallisation Dissolve a volume of the injection containing 50 mg of
from ethanol (95 per cent) melts at about 182° (2.4.21). Nandrolone Phenylpropionate in 8 ml of light petroleum
(40° to 60°) and extract with three 8-ml quantities of a mixture
Tests of 7 volumes of glacial acetic acid and 3 volumes of water.
Wash the combined extracts with 10 ml of light petroleum
Specific optical rotation (2.4.22). +48.0 ° to +51.0°, determined (40° to 60°), dilute with water until the solution becomes
in a 1.0 per cent w/v solution in dioxan. turbid, allow to stand for 2 hours in ice and filter. The precipitate,
Related substances. Determine by thin-layer chromatography after washing with water and drying over phosphorus
(2.4.17), coating the plate with silica gel GF254. pentoxide at a pressure not exceeding 0.7 kPa, complies with
the following test.
Mobile phase. A mixture of 70 volumes of heptane and
30 volumes of acetone. Determine by thin-layer chromatography (2.4.17), using a
silica gel GF254 precoated plate the surface of which has
Test solution. Dissolve 0.1 g of the substance under
been modified by chemically-bonded octadecylsilyl groups.
examination in 100 ml of chloroform.
Mobile phase. A mixture of 20 volumes of water, 40 volumes
Reference solution (a). A 0.005 per cent w/v solution of the of acetonitrile and 60 volumes of propan-2-ol.
substance under examination in chloroform.
Test solution. A 0.5 per cent w/v solution of the dried
Reference solution (b). A 0.01 per cent w/v solution of
precipitate in chloroform.
nandrolone RS in chloroform.
Reference solution (a). A 0.5 per cent w/v solution of
Apply to the plate 5 µl of each solution. After development,
nandrolone phenylpropionate RS in chloroform.
dry the plate in air and examine in ultraviolet light at 254 nm. In
the chromatogram obtained with the test solution any spot Reference solution (b). A mixture of equal volumes of the test
corresponding to nandrolone is not more intense than the solution and the reference solution.
spot in the chromatogram obtained with reference solution Apply to the plate 5 µl of each solution. After development,
(b) and any other secondary spot is not more intense than the dry the plate in air until the solvent has evaporated and heat it
spot in the chromatogram obtained with reference solution at 100° for 10 minutes. Allow to cool and examine in ultraviolet
(a). light at 254 nm. The principal spot in the chromatogram
Sulphated ash (2.3.18). Not more than 0.1 per cent. obtained with the test solution corresponds to that in the
chromatogram obtained with reference solution (a). The
Loss on drying (2.4.19) Not more than 0.5 per cent, determined
principal spot in the chromatogram obtained with reference
on 1.0 g by drying over phosphorus pentoxide at a pressure
solution (b) appears as a single spot.
not exceeding 0.7 kPa for 4 hours.
Assay. Weigh accurately about 10 mg, dissolve in sufficient Tests
ethanol to produce 100.0 ml, dilute 5.0 ml to 50.0 ml with ethanol
Other tests. Complies with the tests stated under Parenteral
and measure the absorbance of the resulting solution at the
Preparations (Injections).
maximum at about 240 nm (2.4.7). Calculate the content of
C27H34O3 taking 430 as the specific absorbance at 240 nm. Assay. To an accurately measured volume containing about
0.1 g of Nandrolone Phenylpropionate add sufficient
Storage. Store protected from light.
chloroform to produce 100.0 ml. Dilute 3.0 ml of this solution
to 50.0 ml with chloroform. To 5.0 ml of the resulting solution
add 10 ml of isoniazid solution and sufficient methanol to
produce 20.0 ml. Allow to stand for 45 minutes and measure
Nandrolone Phenylpropionate the absorbance of the solution at the maximum at about
Injection 380 nm (2.4.7), using as blank 5 ml of chloroform treated in the
same manner. Calculate the content of C27H34O3 from the
Nandrolone Phenylpropionate Injection is a sterile solution
absorbance obtained from a 0.006 per cent w/v solution of
of Nandrolone Phenylpropionate in Ethyl Oleate or other
nandrolone phenylpropionate RS treated in the same manner.
suitable ester, in a suitable fixed oil or in a mixture of these.
Storage. Store protected from light.
Nandrolone Phenylpropionate Injection contains not less than
92.5 per cent and not more than 107.5 per cent of the stated Labelling. The label states that the preparation is for
amount of nandrolone phenylpropionate, C27H34O3. intramuscular injection only.

809
NAPHAZOLINE NITRATE IP 2007

Naphazoline Nitrate Test solution. Dissolve 0.2 g of the substance under


examination in 10 ml of methanol.
N Reference solution. A solution containing 2 per cent w/v of
naphazoline nitrate RS and 0.01 per cent w/v of
N
naphthylacetylethylenediamine hydrochloride RS.
H
, HNO3 Apply to the plate 10 µl of each solution. After development,
dry the plate at 105° for 5 minutes, spray with a 0.5 per cent
w/v solution of ninhydrin in methanol and heat at 105° for
C4HI4N2,HNO3 Mol. Wt. 273.3 10 minutes. Any spot corresponding to naphthylacetyl-
ethylenediamine hydrochloride in the chromatogram obtained
Naphazoline Nitrate is 2-(1-napthylmethyl)-2-imidazoline
with the test solution is not more intense than the
nitrate.
corresponding spot in the chromatogram obtained with the
Naphazoline Nitrate contains not less than 99.0 per cent and reference solution. The test is not valid unless the
not more than 101.0 per cent of C4HI4N2,HNO3 calculated on chromatogram obtained with the reference solution shows
the dried basis. two clearly separated spots.
Description. A white or almost white. crystalline powder. Chlorides (2.3.12). 15.0 ml of 1.0 per cent w/v solution in carbon
dioxide-free water complies with the limit test for chlorides
Identification (375 ppm).
Test A may be omitted if tests B, C and D are carried out. Tests Sulphated ash (2.3.18). Not more than 0.1 per cent.
B and C may be omitted if tests A and D are carried out.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
A. Determine by infrared absorption spectrophotometry (2.4.6). on 1.0 g by drying in an oven at 105° for 3 hours.
Compare the spectrum with that obtained with naphazoline
Assay. Weigh accurately about 0.2 g, dissolve in 30 ml of
nitrate RS.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
B. When examined in the range 230 nm to 360 nm (2.4.7), a acid, determining the end-point potentiometrically (2.4.25).
0.002 per cent w/v solution in 0.01 M hydrochloric acid shows Carry out a blank titration.
absorption maxima at about 270 nm, 280 nm, 287 nm and
1 ml of 0.1 M perchloric acid is equivalent to 0.02733 g of
291 nm; absorbances at these maxima are about 0.43, 0.50, 0.35
C4HI4N2,HNO3.
and 0.34 respectively.
Storage. Store protected from light.
C. Dissolve about 0.5 mg in 1 m1 of methanol, add 0.5 ml of a
freshly prepared 5 per cent w/v solution of sodium
nitroprusside and 0.5 ml of a 2 per cent w/v solution of sodium
hydroxide, allow to stand for 10 minutes and add 1 ml of a
8 per cent w/v solution of sodium bicarbonate; a violet colour Nelfinavir Mesylate
is produced.
D. Dissolve about 10 mg in 5 ml of water, add 0.2 g of magnesium
oxide, shake mechanically for 30 minutes. add 10 ml of
chloroform and shake vigorously. Allow to stand, separate H
CH3
S O N
the chloroform layer, filter and evaporate the aqueous layer to CH3 O CH3
dryness. The residue gives reaction A for nitrates (2.3.1). HO CH3
N N , CH3SO3H
H H
Tests OH
H
Appearance of solution. A 1.0 per cent w/v solution in carbon
dioxide-free water is clear (2.4.1) and colourless (2.4.1).
pH (2.4.24). 5.0 to 6.5, determined in a 1.0 per cent w/v solution. C32H45N3O4S,CH4O3S Mol. Wt. 663.9
Naphthylacetylethylenediamine. Determine by thin-layer Nelfinavir Mesylate is (3S,4aS,8aS)-N-(tert-butyldecahydro-
chromatography (2.4.17), coating the plate with silica gel G. 2-[(2R,3R)-3-(3-hydroxy-o-toluamido)-hydroxy-4-
Mobile phase. A mixture of 100 volumes of methanol and (phenylthio)butyl]isoquinoline-3-carboxamide methyl
1.5 volumes of strong ammonia solution. sulphonate.

810
IP 2007 NELFINAVIR MESTYLATE ORAL POWDER

Nelfinavir Mesylate contains not less than 98.0 per cent and 1 ml of 0.1 M sodium hydroxide is equivalent to 0.00961 g of
not more than 101.0 per cent of C32H45N3O 4S,CH4O3S, CH3SO3H.
calculated on the anhydrous basis.
Heavy metals (2.3.13). 1.0 g complies with the limit test for
Description. A white or almost white powder. heavy metals, Method B (20 ppm).
Identification Sulphated ash (2.3.18). Not more than 0.1 per cent.

A. Determine by infrared absorption spectrophotometry (2.4.6). Water (2.3.43). Not more than 3.0 per cent, determined on
Compare the spectrum with that obtained with nelfinavir 0.5 g.
mesylate RS or with the reference spectrum of nelfinavir Assay. Determine by liquid chromatography (2.4.14).
mesylate.
Test solution. A 0.01 per cent w/v solution of the substance
B. In the Assay, the principal peak in the chromatogram under examination in the mobile phase.
obtained with the test solution corresponds to the peak in the
chromatogram obtained with the reference solution. Reference solution. A 0.01 per cent w/v solution of nelfinavir
mesylate RS in the mobile phase.
Tests
Chromatographic system
Specific optical rotation (2.4.22). –105° to –120°, determined – a stainless steel column 25 cm x 4.6 mm, packed with
in a 1.0 per cent w/v solution in methanol. octadecylsilane bonded to porous silica (5 µm),
Related substances. Determine by liquid chromatography – mobile phase: a filtered and degassed mixture of
(2.4.14), using the chromatographic system described in the 45 volumes of acetonitrile, 20 volumes of methanol
Assay. and 35 volumes of a buffer prepared by dissolving 4.0 g
of sodium dihydrogen phosphate in 1000 ml of water,
Test solution. A 0.1 per cent w/v solution of the substance
to which 1 ml of dimethylamine solution and 1 g of
under examination in the mobile phase.
sodium octanesulphonate are added and mixed to
Reference solution (a). A 0.001 per cent w/v solution of the dissolve,
substance under examination in the mobile phase. – flow rate. 1 ml per minute,
Reference solution (b). A 0.01 per cent w/v solution of – spectrophotometer set at 215 nm,
methanesulphonic acid in the mobile phase. – a 20 µl loop injector.
Inject reference solution (a). The test is not valid unless the Inject the reference solution. The test is not valid unless the
column efficiency determined from the nelfinavir peak is not column efficiency determined from the nelfinavir peak is not
less than 4000 theoretical plates and the tailing factor is not less than 5000 theoretical plates, the tailing factor is not more
more than 2.0. than 2.0 and the relative standard deviation for replicate
Separately inject reference solution (b) and record the injections is not more than 2.0 per cent.
chromatograms. Separately inject the test solution and Separately inject the test solution and the reference solution
continue the chromatography for at least three times the and measure the responses for the principal peak. Calculate
retention time of the principal peak. In the chromatogram the content of C32H45N3O4S,CH4O3S.
obtained with the test solution, the area of any peak other
than the principal peak is not greater than half of the area of Storage. Store protected from light.
the principal peak in the chromatogram obtained with reference
solution (a) (0.5 per cent) and the sum of the areas of all such
peaks is not greater than the area of the principal peak in the
chromatogram obtained with reference solution (a) (1.0 per Nelfinavir Mesylate Oral Powder
cent). Ignore any peak due to methanesulphonic acid
corresponding to the retention time of the principal peak in Nelfinavir Mesylate Oral Powder contains not less than
the chromatogram obtained with reference solution (b). 90.0 per cent and not more than 110.0 per cent of the stated
amount of nelfinavir, C32H45N3O4S.
Methanesulphonic acid. 13.5 per cent to 15.5 per cent w/w,
calculated on the anhydrous basis, determined by the following Identification
method. Weigh accurately about 0.6 g, dissolve in 50 ml of
dimethylformamide and titrate with 0.1 M sodium hydroxide, In the Assay, the principal peak in the chromatogram obtained
determining the end-point potentiometrically (2.4.25). Carry with the test solution corresponds to the peak in the
out a blank titration. chromatogram obtained with the reference solution.

811
NELFINAVIR TABLETS IP 2007

Tests (1.0 per cent) and the sum of areas of all the secondary peaks
is not more than twice the area of the peak in the chromatogram
Dissolution (2.5.2). obtained with the reference solution (b) (2.0 per cent).
Apparatus. No 1
Water (2.3.43). Not more than 12.0 per cent, determined on 0.5 g.
Medium. 900 ml of 0.1 M hydrochloric acid.
Speed and time. 75 rpm and 45 minutes. Assay. Determine by liquid chromatography (2.4.14).
Solvent mixture. 30 volumes of water and 70 volumes of
Withdraw a suitable volume of the medium and filter.
methanol.
Determine by liquid chromatography (2.4.14).
Test solution. Weigh accurately a quantity of the powder
Test solution. Use the filtrate and, if necessary, dilute with the containing 50 mg of Nelfinavir Mestlate, disperse in 50 ml of
dissolution medium. 0.1 M hydrochloric acid, dilute to 250.0 ml with the solvent
Reference solution. A 0.065 per cent w/v solution of nelfinavir mixture and filter.
mesylate RS in methanol. Dilute 10 ml of the solution to 100 ml Reference solution. Dissolve 10 mg of nelfinavir mesylate RS
with the dissolution medium. in 10 ml of 0.1 M hydrochloric acid and dilute to 50.0 ml with
Use the chromatographic system described under Assay. the solvent mixture.
Inject the test solution and the reference solution. Chromatographic system
– a stainless steel column 15 cm x 4.6 mm, packed with
D. Not less than 75 per cent of the stated amount of octadecylsilane bonded to porous silica (5 µm),
C32H45N3O4S. – column temperature 40º,
Related substances. Determine by liquid chromatography – mobile phase: a mixture of 35 volumes of a buffer solution
(2.4.14). prepared by dissolving 4 g of sodium dihydrogen
Test solution. Weigh accurately a quantity of the oral powder phosphate dihydrate and 1g of 1-octane sulphonic
containing 50 mg of Nelfinavir Mesylate, disperse in 10 ml of acid sodium salt into 1000 ml of water, adding 1ml of
methanol, dilute to 50 ml with the mobile phase and filter. dimethylamine and filtering, 45 volumes acetonitrile
and 20 volumes of methanol,
Reference solution (a). Dissolve 10 mg of nelfinavir mesylate – flow rate. 2 ml per minute,
RS in 2 ml of methanol and dilute to 10 ml with the mobile – spectrophotometer set at 220 nm,
phase. – a 10 µl loop injector.
Reference solution (b). Dilute 1 ml of reference solution (a) to Inject the reference solution. The test is not valid unless the
100 ml with the mobile phase. tailing factor is not more than 2.0, the column efficiency in not
Chromatographic system less than 2000 theoretical plates and the relative standard
– a stainless steel column 15 cm x 4.6 mm, packed with deviation for replicate injections is not more than 2.0 per cent.
octadecylsilane bonded to porous silica (5 µm), Inject the test solution and the reference solution.
– column temperature 45º,
– mobile phase: a mixture of 28 volumes of a buffer solution Calculate the content of C32H45N3O4S in the oral powder.
prepared by dissolving 4.88 g of anhydrous sodium Storage. Store protected from moisture, at a temperature not
dihydrogen phosphate in 1000 ml of water, adjusting exceeding 30º.
the pH to 3.4 with phosphoric acid and filtering, Labelling. The label states the strength in terms of the
27 volumes of acetonitrile, 20 volumes of methanol equivalent amount of nelfinavir.
and 25 volumes of water. Adjust the pH to 4.8 with 0.1
M sodium hydroxide or orthophosphoric acid.
– flow rate. 1 ml per minute,
– spectrophotometer set at 220 nm,
Nelfinavir Tablets
– a 10 µl loop injector. Nelfinavir Mesylate Tablets
Inject the reference solution (a). The test is not valid unless Nelfinavir Tablets contain not less than 90.0 per cent and not
the tailing factor is not more than 2.0 and the column efficiency more than 110.0 per cent of the stated amount of nelfinavir
in not less than 4000 theoretical plates. mesylate, C32H45N3O4S,CH4O3S.
Inject the test solution and reference solution (b). In the
Identification
chromatogram obtained with the test solution, the area of any
secondary peak is not more than the area of the peak in the A. Shake a quantity of the powdered tablets containing about
chromatogram obtained with the reference solution (b) 0.1 g of Nelfinavir Mesylate with 80 ml of methanol for

812
IP 2007 NEOMYCIN SULPHATE

10 minutes, add sufficient methanol to produce 100 ml, mix Inject separately the diluent (10 ml of methanol diluted to
and filter. Dilute 5 ml of the filtrate to 100 ml with methanol. 50 ml with the mobile phase) and the test solution and continue
When examined in the range 200 nm to 300 nm the resulting the chromatography for 4 times the retention time of the
solution shows an absorption maximum only at about 254 nm principal peak. Examine the diluent chromatogram for any
(2.4.7). extraneous peaks and ignore the corresponding peaks
observed in the chromatogram obtained with the test solution.
B. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak in the Any secondary peak observed in the chromatogram obtained
chromatogram obtained with the reference solution. with the test solution should not be more than 1.0 per cent
and the sum of the areas of all the secondary peaks should
Tests not be more than 2.0 per cent when calculated by percentage
Dissolution (2.5.2). area normalisation. Inhibit integration of peak due to
Apparatus. No 1 methanesulphonic acid.
Medium. 900 ml of 0.01 M hydrochloric acid. Other tests. Complies with the tests stated under Tablets.
Speed and time. 50 rpm and 30 minutes. Assay. Determine by liquid chromatography (2.4.14).
Withdraw a suitable volume of the medium and filter promptly Test solution. Weigh accurately a quantity of the powdered
through a membrane filter disc with an average pore diameter tablets containing about 200 mg of Nelfinavir Mesylate, add
not greater than 1.0 µm. Reject the first few ml of the filtrate about 20 ml of methanol, mix with the aid of ultrasound for
and dilute a suitable volume of the filtrate with the same 10 minutes and dilute to 100.0 ml with the mobile phase. Filter
solvent. Measure the absorbance of the resulting solution at through a membrane filter disc with an average pore diameter
the maximum at about 250 nm (2.4.7). Calculate the content of not greater than 1.0 µm, rejecting the first few ml of the filtrate.
C32H45N3O4S,CH4O3S from the absorbance of a solution of Further dilute 5.0 ml of the filtrate to 100.0 ml with the mobile
known concentration of nelfinavir mesylate RS. phase.
D. Not less than 75 per cent of the stated amount of Reference solution. Weigh accurately about 50 mg of
C32H45N3O4S, CH4O3S. nelfinavir mesylate RS, add about 10 ml of methanol, mix with
Related substances. Determine by liquid chromatography the aid of ultrasound to dissolve and dilute to 50.0 ml with the
(2.4.14). mobile phase. Dilute 5.0 ml of this solution to 50.0 ml with the
Test solution. Weigh accurately a quantity of the powdered mobile phase.
tablets containing about 100 mg of Nelfinavir Mesylate, add Use the chromatographic system described in the test for
about 20 ml of methanol, mix with the aid of ultrasound for Related substances.
10 minutes and dilute to 100 ml with the mobile phase. Inject the reference solution. The test is not valid unless the
Reference solution. Weigh accurately about 10 mg of column efficiency determined from the nelfinavir mesylate peak
nelfinavir mesylate RS, add about 10 ml of methanol, shake is not less than 5000 theoretical plates, the tailing factor is not
for 10 minutes and dilute to 50 ml with the mobile phase. more than 2.0 and the relative standard deviation for replicate
Chromatographic system injections is not more than 2.0 per cent.
– a stainless steel column 25 cm x 4.6 mm, packed with Inject separately the test solution and the reference solution
octadecylsilane bonded to porous silica particles or and measure the responses for the major peak. Calculate the
ceramic microparticles (5 µm), content of C32H45N3O4S,CH4O3S in the tablets.
– mobile phase: a filtered and degassed mixture of
Storage. Store protected from light.
45 volumes of acetonitrile, 20 volumes of methanol
and 35 volumes of a buffer prepared by dissolving 4.0 g
of sodium dihydrogen phosphate in 1000 ml of water,
to which are added 1 ml of dimethylamine solution and Neomycin Sulphate
1 g of sodium octanesulphonate and mixing to dissolve, Neomycin Sulphate is a mixture of the sulphates of substances
– flow rate. 1 ml per minute, obtained by the growth of certain selected strains of
– spectrophotometer set at 215 nm, Streptomyces fradiae.
– a 20 µl loop injector.
Neomycin Sulphate has a potency of not less than 600 Units
Inject the reference solution. The test is not valid unless the
per mg, calculated on the dried basis.
column efficiency determined from the nelfinavir mesylate peak
is not less than 4000 theoretical plates and the tailing factor is Description. A white or yellowish-white powder; odourless
not more than 2.0. or almost odourless; hygroscopic.

813
NEOMYCIN EYE DROPS IP 2007

Identification Mobile phase. A mixture of 80 volumes of a 20 per cent w/v


solution of sodium chloride and 20 volumes of methanol.
A. Determine by thin-layer chromatography (2.4.17), coating
the plate with silica gel H. Test solution. Dissolve 40 mg of the substance under
examination in water and dilute to 5 ml with the same solvent.
Mobile phase. A freshly prepared 3.85 per cent w/v solution
of ammonium acetate. Reference solution (a). Dissolve 30 mg of framycetin sulphate
RS in water and dilute to 25 ml with the same solvent.
Test solution. Dissolve 0.2 g of the substance under
Reference solution (b). Dilute 5 ml of reference solution (a) to
examination in 10 ml of water.
25 ml with water.
Reference solution. A 2.0 per cent w/v solution of neomycin
Reference solution (c). Dissolve 40 mg of neomycin sulphate
sulphate RS in water.
RS in water and dilute to 5 ml with the same solvent.
Apply to the plate 1 µl of each solution. After development,
Apply to the plate as 5-mm bands 5 µl of each solution. Dry
dry the plate in air for 10 minutes, heat at 100° for 1 hour and
the bands; allow the mobile phase to rise at least 12 cm. Dry
spray with a 0.1 per cent w/v solution of ninhydrin in
the plate at 100° to 105° for 10 minutes. Spray the plate with
1-butanol saturated with water. Heat again at 100° for
ethanolic ninhydrin solution and heat at 100° to 105° for
5 minutes. The principal spot in the chromatogram obtained
10 minutes. In the chromatogram obtained with the test
with the test solution corresponds to that in the chromatogram
solution the principal band corresponds to the principal band
obtained with the reference solution.
in the chromatogram obtained with reference solution (c) and
B. Dissolve about 10 mg in 5 ml of water, add 0.1 ml of pyridine the band due to neomycin C with an Rf value slightly less than
and 2 ml of a 0.1 per cent w/v solution of ninhydrin and heat that of the principal band is not more intense than the band
on a water-bath at a temperature of about 70° for 10 minutes; obtained with reference solution (a) (15 per cent) but is more
a deep violet colour is produced. intense than the band in the chromatogram obtained with
C. A 5 per cent w/v solution gives the reactions of sulphates reference solution (b) (3 per cent). The test is not valid unless
(2.3.1). in the chromatogram obtained with reference solution (c) a
band appears with an Rf value slightly less than that of the
Tests principal band.
pH (2.4.24). 5.0 to 7.5, determined in a 1.0 per cent w/v solution. Sulphated ash (2.3.18). Not more than 1.0 per cent.
Specific optical rotation (2.4.22).+53.5° to +59.0°, determined Loss on drying (2.4.19). Not more than 8.0 per cent, determined
in a 10.0 per cent w/v solution. on 0.5 g by drying in an oven at 60° over phosphorus pentoxide
at a pressure not exceeding 0.7 kPa for 3 hours.
Neamine. Determine by thin-layer chromatography (2.4.17),
coating the plate with silica gel H. Assay. Determine by the microbiological assay of antibiotics,
Method A (2.2.10).
Mbile phase. A mixture of 30 volumes of methanol, 20 volumes
of strong ammonia solution and 10 volumes of Storage. Store protected from light and moisture.
dichloromethane. Labelling. The label states the strength in terms of Units of
Test solution. Dissolve 0.25 g of the substance under neomycin per mg.
examination in 10 ml of water.
Reference solution. A 0.05 per cent w/v solution of neamine
RS in water. Neomycin Eye Drops
Apply to the plate as 5-mm bands 5 µl of each solution. Dry Neomycin Sulphate Eye Drops
the bands; allow the mobile phase to rise at least 8 cm. Dry the
Neomycin Sulphate Eye Drops are a sterile solution of
plate in a current of warm air, heat at 110° for 10 minutes, spray
Neomycin Sulphate in Purified Water.
the plate with ninhydrin and stannous chloride reagent and
heat at 110° for 15 minutes. Spray the plate again with the Neomycin Sulphate Eye Drops contain not less than 90.0 per
same reagent and heat at 110° for 15 minutes. Any band cent and not more than 115.0 per cent w/v of the stated amount
corresponding to neamine in the chromatogram obtained with of neomycin sulphate.
the test solution is not more intense than the spot in the
chromatogram obtained with the reference solution. Identification
Neomycin C. Determine by thin-layer chromatography (2.4.17), Determine by thin-layer chromatography (2.4.17), coating the
coating the plate with silica gel of a suitable grade. plate with silica gel.

814
IP 2007 NEOMYCIN EYE OINTMENT

Mobile phase. A mixture of 60 volumes of methanol, Chromatographic system


40 volumes of strong ammonia solution and 20 volumes of – a stainless steel column 20 cm x 4.6 mm, packed with
chloroform. porous silica particles (5 µm) (such as Nucleosil 100-5),
Test solution. Dilute if necessary a volume of the eye drops to – mobile phase: a mixture of 97 ml of tetrahydrofuran,
produce a solution containing 0.5 per cent w/v of Neomycin 1.0 ml of water and 0.5 ml of glacial acetic acid diluted
Sulphate in water. with sufficient of a 2.0 per cent v/v solution of ethanol
in ethanol-free chloroform to produce 250 ml,
Reference solution (a). A 0.5 per cent w/v solution of – flow rate. 1.6 ml per minute,
neomycin sulphate RS in water. – spectrophotometer set at 350 nm,
Reference solution (b). A mixture of equal volumes of the eye – a 10 µl loop injector.
drops and reference solution (a). If necessary the tetrahydrofuran and water content of the
Apply to the plate 1 µl of each solution. After development, mobile phase may be adjusted so that the chromatogram
dry the plate in air, spray with a 1 per cent w/v solution of obtained with the reference solution shows resolution similar
ninhydrin in 1-butanol and heat at 105° for 2 minutes. The to that in the specimen chromatogram supplied with framycetin
principal red spot in the chromatogram obtained with the test sulphate RS. The mobile phase should be passed through the
solution corresponds to that in the chromatogram obtained column for several hours before the solutions are injected.
with reference solution (a) and the principal red spot in the Continue the chromatography for 1.4 times the retention time
chromatogram obtained with reference solution (b) appears of the peak due to neomycin B.
as a single spot. The column efficiency, determined using the peak due to
Neomycin B in the chromatogram obtained with the test
Tests solution, should be not less than 13,000 theoretical plates.
Neamine. Determine by thin-layer chromatography (2.4.17), In the chromatogram obtained with the test solution the area
coating the plate with silica gel H. of the peak corresponding to neomycin C is not less than
Mobile phase. A mixture of 30 volumes of methanol, 3.0 per cent and not more than 15.0 per cent of sum of the areas
20 volumes of strong ammonia solution and 10 volumes of of the peaks corresponding to Neomycin B and Neomycin C.
dichloromethane. Other Tests. Complies with the tests stated under Eye Drops.
Test solution. A volume of the eye drops containing 5 µg Assay. Measure accurately a quantity containing 5 mg of
(3.5 Units). Neomycin Sulphate and dilute to 50.0 ml with sterile phosphate
Reference solution. The same volume of water containing buffer pH 8.0 and mix. Dilute 10.0 ml of the resulting solution
0.1 µg of neamine RS. to 100.0 ml with the same solvent.

Apply to the plate each solution. After development, dry the Determine by the microbiological assay of antibiotics, Method
plate in a current of warm air, heat at 110° for 10 minutes, spray A (2.2.10)
the plate with ninhydrin and stannous chloride reagent and The upper fiducial limit of error is not less than 90.0 per cent
heat at 110° for 15 minutes. Spray the plate again with the and the lower fiducial limit of error is not more than 115.0 per
same reagent and heat at 110° for 15 minutes. Any spot cent of the stated number of Units per ml.
corresponding to neamine in the chromatogram obtained with
Storage. Store protected from light.
the test solution is not more intense than the spot in the
chromatogram obtained with the reference solution. Labelling. The strength is stated in terms of percentage w/v
as well as the number of Units per ml.
Neomycin C. Determine by liquid chromatography (2.4.14).
Test solution. Dilute the eye drops with 0.02 M borax to
contain 1 mg (700 Units) per ml. To 0.5 ml of the diluted solution
add 1.5 ml of a freshly prepared 2 per cent w/v solution of Neomycin Eye Ointment
1-fluoro-2,4-dinitrobenzene in methanol, dilute to 25 ml with
Neomycin Sulphate Eye Ointment
the mobile phase, allow to stand and use the clear lower layer.
Neomycin Sulphate Eye Ointment is a sterile preparation
Reference solution. Add 1.5 ml of the 1-fluoro-2,4-
containing Neomycin Sulphate in a suitable basis.
dinitrobenzene solution to 0.5 ml of a 0.1 per cent w/v solution
of neomycin sulphate RS in 0.02 M borax, heat in a water- Neomycin Sulphate Eye Ointment contains not less than
bath at 60° for 1 hour and cool; dilute the solution to 25 ml with 90.0 per cent and not more than 115.0 per cent of the stated
the mobile phase, allow to stand and use the clear lower layer. amount of neomycin sulphate.

815
NEOMYCIN EYE OINTMENT IP 2007

Identification cent w/v solution of 1-fluoro-2,4-dinitrobenzene in methanol,


heat on a water-bath at 60° for 1 hour and cool. Dilute the
Determine by thin-layer chromatography (2.4.17), coating the resulting solution to 25 ml with the mobile phase, allow to
plate with silica gel. stand and use the clear lower layer.
Mobile phase. A mixture of 60 volumes of methanol, 40 volumes Reference solution. Add 1.5 ml of the 1-fluoro-2,4-
of strong ammonia solution and 20 volumes of chloroform. dinitrobenzene solution to 0.5 ml of a 0.1 per cent w/v solution
Test solution. Disperse a quantity of the eye ointment of neomycin sulphate RS in 0.02 M borax and proceed as for
containing 20 mg of Neomycin Sulphate in 20 ml of chloroform, the test solution.
extract with 5 ml of water and use the aqueous extract. Chromatographic system
Reference solution (a). A 0.4 per cent w/v solution of – a stainless steel column 20 cm x 4.6 mm, packed with
neomycin sulphate RS in water. porous silica particles (5 µm),
– mobile phase: 97 ml of tetrahydrofuran, 1.0 ml of water
Reference solution (b). A mixture of equal volumes of test and 0.5 ml of glacial acetic acid with sufficient of a
solution and reference solution (a). 2.0 per cent v/v solution of ethanol in ethanol-free
Apply to the plate 1 µl of each solution. After development, chloroform to produce 250 ml,
dry the plate in air, spray with a 1 per cent w/v solution of – flow rate. 1.6 ml per minute,
ninhydrin in 1-butanol and heat at 105° for 2 minutes. The – spectrophotometer set at 350 nm,
principal red spot in the chromatogram obtained with the test – a 10 µl loop injector.
solution corresponds to that in the chromatogram obtained If necessary the tetrahydrofuran and water content of the
with reference solution (a) and the principal red spot in the mobile phase may be adjusted so that the chromatogram
chromatogram obtained with reference solution (b) appears obtained with reference solution shows resolution similar to
as a single spot. that in the specimen chromatogram supplied with framycetin
sulphate RS. The mobile phase should be passed through the
Tests column for several hours before the solutions are injected.
Neamine. Determine by thin-layer chromatography (2.4.17), Continue the chromatography for 1.4 times the retention time
coating the plate with silica gel H. of the peak due to neomycin B.

Mobile phase. A mixture of 30 volumes of methanol, The column efficiency, determined using the peak due to
20 volumes of strong ammonia solution and 10 volumes of Neomycin B in the chromatogram obtained with the test
dichloromethane. solution, should be not less than 13,000 theoretical plates.
In the chromatogram obtained with the test solution the area
Test solution. Disperse a quantity of the eye ointment
of the peak corresponding to neomycin C is not less than
containing 20 mg of Neomycin Sulphate in 20 ml of chloroform,
3.0 per cent and not more than 15.0 per cent of the sum of the
shake gently with 8 ml of water, allow the layers to separate
areas of the peaks corresponding to Neomycin B and Neomycin
and use the aqueous layer.
C.
Reference solution. A 0.005 per cent w/v solution of neamine
Other Tests. Complies with the tests stated under Eye
RS in water.
Ointments.
Apply to the plate 2 µl of each solution. After development,
Assay. Weigh accurately a quantity containing 5 mg of
dry the plate in a current of warm air, heat at 110° for 10 minutes,
Neomycin Sulphate, dissolve in 25 ml of chloroform, extract
spray with ninhydrin and stannous chloride reagent and
with four quantities, each of 20 ml, of sterile phosphate buffer
heat at 110° for 15 minutes. Spray the plate again with the
pH 8.0, combine the extracts and add sufficient of the buffer
same reagent and heat at 110° for 15 minutes. Any spot
solution to produce 100.0 ml.
corresponding to neamine in the chromatogram obtained with
the test solution is not more intense than the spot in the Carry out the microbiological assay of antibiotics, Method A
chromatogram obtained with the reference solution. (2.2.10).
Neomycin C. Determine by liquid chromatography (2.4.17) The upper fiducial limit of error is not less than 90.0 per cent
and the lower fiducial limit of error is not more than 115.0 per
Test solution. Disperse a quantity of the eye ointment cent of the stated number of Units per g.
containing 5 mg of Neomycin Sulphate in 20 ml of light
petroleum (120° to 160°), add 5 ml of 0.02 M borax, shake, Storage. Store protected from light.
separate the aqueous layer and centrifuge. To 0.5 ml of the Labelling. The strength is stated in terms of percentage w/v
separated aqueous layer add 1.5 ml of a freshly prepared 2 per as well as the number of Units per ml.

816
IP 2007 NEOSTIGMINE TABLETS

Neostigmine Bromide measured immediately after preparation, not more than


0.25 (2.4.7).

CH3 Sulphates (2.3.17). 0.75 g complies with the limit test for
sulphates (200 ppm).
H3C N CH3
Sulphated ash (2.3.18). Not more than 0.1 per cent.
O Br Loss on drying (2.4.19). Not more than 1.0 per cent, determined
CH3 on 1.0 g by drying in an oven at 105°.
O N
CH3 Assay. Weigh accurately about 0.5 g, dissolve in 20 ml of
anhydrous glacial acetic acid, add 5 ml of acetic anhydride.
Titrate with 0.1 M perchloric acid, using crystal violet
C12H19BrN2O2 Mol. Wt. 303.2
solution as indicator. Carry out a blank titration.
Neostigmine Bromide is 3-(dimethylcarbamoyloxy)
1 ml of 0.1 M perchloric acid is equivalent to 0.03032 g of
trimethylanilinium bromide.
C12H19BrN2O2.
Neostigmine Bromide contains not less than 98.0 per cent and
Storage. Store protected from light and moisture.
not more than 101.0 per cent of C12H19BrN2O2, calculated on
the dried basis.
Description. Colourless crystals or a white, crystalline powder;
odourless; hygroscopic.
Neostigmine Tablets
Neostigmine Bromide Tablets
Identification
Neostigmine Bromide Tablets contain not less than 92.5 per
Test A may be omitted if tests B, C, D and E are carried out. cent and not more than 107.5 per cent of the stated amount of
Tests B, C and D may be omitted if tests A and E are carried neostigmine bromide, C12H19BrN2O2.
out.
A. Determine by infrared absorption spectrophotometry (2.4.6). Identification
Compare the spectrum with that obtained with neostigmine Triturate a quantity of the powdered tablets containing about
bromide RS. 0.3 g of Neostigmine Bromide with three quantities, each of
B. When examined in the range 230 nm to 360 nm (2.4.7), a 5 ml of ether and discard the ether. Macerate the residue with
0.02 per cent w/v solution in 0.5 M sulphuric acid shows several quantities, each of 10 ml of ethanol (95 per cent),
absorption maxima at about 260 nm and 266 nm. filtering after each maceration. Evaporate the combined filtrates
on a water-bath and dry the residue at 105° for 1 hour. The
C. Warm about 50 mg with 1 ml of dilute sodium hydroxide
residue melts at about 167°, with decomposition. The residue
solution; an odour of dimethylamine develops slowly.
complies with the following tests.
D. Warm about 50 mg with 0.4 g of potassium hydroxide and
A. Warm about 50 mg with 0.4 g of potassium hydroxide and
2 ml of ethanol (95 per cent) on a water-bath for 3 minutes,
2 ml of ethanol (95 per cent) on a water-bath for 3 minutes,
replacing the evaporated ethanol. Cool, add 2 ml of dilute
replacing the evaporated ethanol. Cool, add 2 ml of dilute
diazobenzenesulphonic acid solution; an orange-red colour
diazobenzenesulphonic acid solution; an orange-red colour
is produced.
is produced.
E. Gives the reactions of bromides (2.3.1). B. Gives the reactions of bromides (2.3.1).
Tests Tests
Appearance of solution. A 0.5 per cent w/v solution is clear
Other Tests. Complies with the tests stated under Tablets.
(2.4.1), and colourless (2.4.1).
Assay. Weigh and powder 20 tablets. Weigh accurately a
Acidity. Dissolve 0.2 g in 20 ml of carbon dioxide-free water
quantity of the powder containing about 0.15 g of Neostigmine
and titrate to pH 7.0 with 0.02 M sodium hydroxide (carbonate-
Bromide, transfer to a semi-micro ammonia-distillation
free); not more than 0.1 ml is required.
apparatus, add 20 ml of a 50 per cent w/v solution of sodium
3-Hydroxytrimethylanilinium bromide. Dissolve 50 mg in a hydroxide and 0.5 ml of a 2 per cent w/v solution of 2-octanol
mixture of 1 ml of sodium carbonate solution and 9 ml of in liquid paraffin. Pass a current of steam through the mixture,
water. Absorbance of the resulting solution at about 294 nm, collect the distillate in 50 ml of 0.01 M sulphuric acid until the

817
NEOSTIGMINE METHYLSULPHATE IP 2007

volume is about 200 ml and titrate the excess of acid with phthalein solution; the solution is colourless. Add
0.02 M sodium hydroxide using methyl red solution as 0.3 ml of 0.01 M sodium hydroxide; the solution becomes red.
indicator. Repeat the operation without the substance under Add 0.4 ml of 0.01 M hydrochloric acid; the solution becomes
examination. The difference between the titrations represents colourless. Add 0.1 ml of methyl red solution; the solution
the amount of sulphuric acid required to neutralise the becomes red or yellowish-red.
dimethylamine produced. 3-Hydroxytrimethylanilinium methyl sulphate. Dissolve
1 ml of 0.01 M sulphuric acid is equivalent to 0.006064 g of 50 mg in a mixture of 1 ml of sodium carbonate solution and
C12H19BrN2O2. 9 ml of water. Absorbance of the resulting solution at about
Storage. Store protected from light and moisture. 294 nm, measured immediately after preparation, not more than
0.20 (2.4.7).
Chlorides (2.3.12). 1.0 g complies with the limit test for
Neostigmine Methylsulphate chlorides (250 ppm).
C13H22N2O6S Mol. Wt. 334.4 Sulphates (2.3.17). 0.75 g complies with the limit test for
sulphates (200 ppm).
Neostigmine Methylsulphate is 3-(dimethylcarbamoyloxy)-
trimethylanilinium methyl sulphate. Loss on drying (2.4.19). Not more than 1.0 per cent, determined
on 1.0 g by drying in an oven at 105°.
Neostigmine Methylsulphate contains not less than 98.5 per
cent and not more than 101.0 per cent of C13H22N2O6S, Sulphated ash (2.3.18). Not more than 0.1 per cent.
calculated on the dried basis. Assay. Weigh accurately about 0.3 g and dissolve in 150 ml of
Description. Colourless crystals or a white, crystalline powder; water. Add 100 ml of 2 M sodium hydroxide, distill and collect
hygroscopic. the distillate in 50 ml of a 4 per cent w/v solution of boric acid
until a total volume of 250 ml is reached. Titrate the distillate
Identification with 0.1 M hydrochloric acid using 0.25 ml of methyl red-
methylene blue solution as indicator. Repeat the operation
Test A may be omitted if tests B, C and D are carried out. Tests
without the substance under examination. The difference
B, C and D may be omitted if test A is carried out.
between the titrations represents the amount of hydrochloric
A. Determine by infrared absorption spectrophotometry (2.4.6). acid required.
Compare the spectrum with that obtained with neostigmine
1 ml of 0.1 M hydrochloric acid is equivalent to 0.03344 g of
methylsulphate RS or with the reference spectrum of
C13H22N2O6S.
neostigmine methylsulphate.
Storage. Store protected from light and moisture.
B. When examined in the range 230 nm to 360 nm (2.4.7), a
0.05 per cent w/v solution in 0.5 M sulphuric acid shows
absorption maxima, at about 261 nm and 267 nm. The ratio of
the absorbance at the maximum at about 267 nm to that at the Neostigmine Injection
maximum at 261 nm is 0.84 to 0.87.
Neostigmine Methylsulphate Injection
C. Dissolve 0.1 g in 5 ml of distilled water and add 1 ml of a
6 per cent w/v solution of barium chloride; no precipitate is Neostigmine Injection is a sterile solution of Neostigmine
produced. Add 2 ml of hydrochloric acid and heat in a water- Methylsulphate in Water for Injections.
bath for 10 minutes; a white precipitate is produced. Neostigmine Injection contains not less than 90.0 per cent
D. Warm about 50 mg with 0.4 g of potassium hydroxide and and not more than 110.0 per cent of the stated amount of
2 ml of ethanol (95 per cent) on a water-bath for 3 minutes, neostigmine methylsulphate, C13H22N2O6S.
replacing the evaporated ethanol. Cool, add 2 ml of dilute
Identification
diazobenzenesulphonic acid solution; an orange-red colour
is produced. A. Dilute, if necessary, a volume of the injection containing
2.5 mg of Neostigmine Methylsulphate to 5 ml with water,
Tests shake with three quantities, each of 10 ml, of ether and discard
Appearance of solution. A 5.0 per cent w/v solution in distilled the ether extracts.
water is clear (2.4.1), and colourless (2.4.1). When examined in the range 230 nm to 360 nm (2.4.7), a 2 cm
Acidity or alkalinity. To 4.0 ml of a 5.0 per cent w/v solution in layer of the resulting solution shows absorption maxima at
distilled water add 6.0 ml of water and 0.1 ml of phenol- about 260 nm and 267 nm.

818
IP 2007 NEVIRAPINE

B. Determine by thin-layer chromatography (2.4.17), coating – spectrophotometer set at 215 nm,


the plate with silica gel G. – a 10 µl loop injector.
Mobile phase. A mixture of 50 volumes of chloroform, In the chromatogram obtained with reference solution (b) the
35 volumes of methanol, 10 volumes of formic acid and principal peak has a retention time of about 6.8 minutes
5 volumes of water. (neostigmine methylsulphate) and there is a peak with a
relative retention time of about 0.5 ((3-hydroxy)
Test solution. Dilute the injection under examination, if
trimethylanilinium methylsulphate). In the chromatogram
necessary, with water to produce a solution containing
obtained with the test solution, the area of any secondary
0.05 per cent w/v of Neostigmine Methylsulphate.
peak with a retention time corresponding to that of the peak
Reference solution (a). A 0.05 per cent w/v solution of due to (3-hydroxy)trimethylanilinium methylsulphate in the
neostigmine methylsulphate RS in water. chromatogram obtained with reference solution (b) is not
Reference solution (b). A mixture of equal volumes of the test greater than the area of the principal peak in the chromatogram
solution and reference solution (a). obtained with reference solution (a) (1 per cent).

Apply to the plate 10 µl of each solution. After development, Other Tests. Complies with the tests stated under Parenteral
dry the plate in air, spray with dilute potassium Preparations (Injections).
iodobismuthate solution. The principal spot in the Assay. Dilute an accurately measured volume containing about
chromatogram obtained with the test solution corresponds to 25 mg of Neostigmine Methylsulphate to 50.0 ml with water.
that in the chromatogram obtained with reference solution (a). Measure the absorbance of the resulting solution at the
The principal spot in the chromatogram obtained with maximum at about 260 nm (2.4.7). Calculate the content of
reference solution (b) appears as a single, compact spot. C13H22N2O6S taking 14.35 as the specific absorbance at
C. To 1 ml add 0.5 ml of sodium hydroxide solution and 260 nm.
evaporate to dryness on a water-bath. Heat quickly in an oil- Storage. Store protected from light.
bath to about 250° and maintain at this temperature for about
30 seconds. Cool, dissolve the residue in 1 ml of water, cool in
ice water and add 1 ml of diazobenzenesulphonic acid
solution; an orange-red colour is produced. Nevirapine
Tests
O
pH. (2.4.24) 4.5 to 6.5. H 3C
HN
3-Hydroxy trimethylanilinium methyl sulphate. Determine
by liquid chromatography (2.4.14).
N N N
Test solution. Dilute the injection if necessary, with water to
contain a 0.05 per cent w/v solution of Neostigmine
Methylsulphate.
Reference solution (a). Dilute 1 volume of the test solution to C15H14N4O Mol. Wt. 266.3
100 volumes with water. Nevirapine is 11-cyclopropyl-5,11-dihydro-4-methy-6H-
Reference solution (b). Add 0.05 ml of 5 M sodium hydroxide dipyrido[3,2-b:2′,3′-e][1,4]diazepin-6-one.
to 1 ml of the test solution and allow to stand for 5 minutes. Nevirapine contains not less than 98.0 per cent and not more
Add 0.1 ml of 5 M hydrochloric acid and use immediately. than 102.0 per cent of C15H14N4O, calculated on the dried basis.
Chromatographic system Description. A white or almost white crystalline powder.
– a stainless steel column 25 cm × 4.6 mm packed with
octadecylsilane chemically bonded to porous silica Identification
particles (5 µm) (such as Lichrosphere 60 RP-select B),
A. Determine by infrared absorption spectrophotometry (2.4.6).
– mobile phase: 0.0015 M solution of sodium
Compare the spectrum with that obtained with nevirapine RS
heptanesulphonate in a mixture of 15 volumes of
or with the reference spectrum of nevirapine.
acetonitrile and 85 volumes of 0.05 M potassium
dihydrogen orthophosphate adjusted to pH 3.0 with B. In the Assay, the principal peak in the chromatogram
orthophosphoric acid, obtained with the test solution corresponds to that in the
– flow rate. of 1.1 ml per minute, chromatogram obtained with the reference solution.

819
NEVIRAPINE TABLETS IP 2007

Tests and 60 volumes of a buffer prepared by dissolving


12.0 g of sodium dihydrogen phosphate in about 800 ml
Related substances. Determine by liquid chromatography of water, adjusting the pH to 3.0 with orthophosphoric
(2.4.14). acid and diluting to 1000.0 ml with water,
Test solution. Dissolve 0.1 g of the substance under – flow rate. 1.2 ml per minute,
examination in 100 ml of methanol. – spectrophotometer set at 230 nm,
– a 20 µl loop injector.
Reference solution. Dilute 1 ml of the test solution to 100 ml
with methanol. Inject the reference solution. The test is not valid unless the
Chromatographic system column efficiency determined from the nevirapine peak is not
– a stainless steel column 25 cm x 4.6 mm, packed with less than 3000 theoretical plates, the tailing factor is not more
octadecylsilane bonded to porous silica or ceramic than 2.0 and the relative standard deviation for the replicate
microparticles (5 µm), injections is not more than 2.0 per cent.
– mobile phase: a filtered and degassed mixture of Separately inject the test solution and the reference solution
20 volumes of methanol, 20 volumes of acetonitrile and measure the responses for the principal peak. Calculate
and 60 volumes of a buffer prepared by dissolving the content of C15H14N4O.
12.0 g of sodium dihydrogen phosphate in about 800 ml
Storage. Store protected from moisture.
of water, adjusting the pH to 3.0 with phosphoric acid
and diluting to 1000.0 ml with water,
– flow rate. 1.2 ml per minute,
– spectrophotometer set at 230 nm, Nevirapine Tablets
– a 20 µl loop injector.
Nevirapine Tablets contain not less than 90.0 per cent and not
Inject the reference solution. The test is not valid unless the more than 110.0 per cent of the stated amount of nevirapine,
column efficiency determined from the nevirapine peak is not C15H14N4O.
less than 5000 theoretical plates and the tailing factor is not
more than 2.0. Identification
Separately inject the test solution and the reference solution. A. When examined in the range 250 nm to 450 nm (2.4.7) a
In the chromatogram obtained with the test solution, the area 0.001 per cent w/v solution in the mobile phase described
of any peak other than the principal peak is not greater than under Assay, shows an absorption maximum at about 230 nm.
half of the area of the principal peak in the chromatogram
B. In the Assay, the principal peak in the chromatogram
obtained with the reference solution (0.5 per cent) and the
obtained with the test solution corresponds to the peak in the
sum of the areas of all such peaks is not greater than the area
chromatogram obtained with the reference solution.
of the principal peak in the chromatogram obtained with the
reference solution (1.0 per cent). Dissolution (2.5.2).
Heavy metals (2.3.13). 1.0 g complies with the limit test for Apparatus. No 1.
heavy metals, Method B (20 ppm). Medium. 900 ml of 0.1 M hydrochloric acid.
Sulphated ash (2.3.18). Not more than 0.2 per cent. Speed and time. 50 rpm and 45 minutes.

Loss on drying (2.4.19). Not more than 0.5 per cent, determined Withdraw a suitable volume of the medium and filter through
on 1.0 g by drying at 105° for 3 hours. a membrane filter disc with an average pore diameter not greater
than 1.0 µm, rejecting the first few ml of the filtrate. Measure
Assay: Determine by liquid chromatography (2.4.14). the absorbance of the resulting solution, at the maximum at
Test solution. A 0.005 per cent w/v solution of the substance about 313 nm (2.4.7).
under examination in methanol.
Calculate the content of C15H14N4O from the absorbance
Reference solution. A 0.005 per cent w/v solution of obtained from a solution of known concentration of nevirapine
nevirapine RS in methanol. RS in 0.1 M hydrochloric acid.
Chromatographic system D. Not less than 70 per cent of the stated amount of C15H14N4O.
– a stainless steel column 25 cm x 4.6 mm, packed with
Related substances. Determine by liquid chromatography
octadecylsilane bonded to porous silica or ceramic
(2.4.14).
microparticles (5 µm),
– mobile phase: a filtered and degassed mixture of Test solution. Shake a quantity of the powdered tablets with a
20 volumes of methanol, 20 volumes of acetonitrile suitable quantity of the mobile phase to obtain a mixture

820
IP 2007 NEVIRAPINE ORAL SUSPENSION

containing 0.05 per cent w/v of Nevirapine and filter through – flow rate. 1.2 ml per minute,
a membrane filter disc with an average diameter not exceeding – spectrophotometer set at 230 nm,
1.0 µm, rejecting the first few ml of the filtrate. – a 20 µl loop injector.
Reference solution. A 0.05 per cent w/v solution of nevirapine Inject the reference solution. The test is not valid unless the
RS in the mobile phase. column efficiency determined from the nevirapine peak is not
less than 7500 theoretical plates, the tailing factor is not more
Chromatographic system
than 1.5 and the relative standard deviation for replicate
– a stainless steel column 25 cm x 4.6 mm, packed with
injections is not more than 2.0 per cent.
octadecylsilane bonded to porous silica or ceramic
microparticles (5 µm), Separately inject the test solution and the reference solution
– mobile phase: a filtered and degassed mixture of and measure the responses for the principal peak. Calculate
20 volumes of methanol, 20 volumes of acetonitrile the content of C15H14N4O in the tablets.
and 60 volumes of a buffer prepared by dissolving
Storage. Store protected from moisture at a temperature not
12.0 g of sodium dihydrogen phosphate in about 800 ml
exceeding 30°.
of water, adjusting the pH to 3.0 with orthophosphoric
acid and diluting to 1000.0 ml with water,
– flow rate. 1.2 ml per minute,
– spectrophotometer set at 230 nm, Nevirapine Oral Suspension
– a 20 µl loop injector.
Nevirapine Oral Suspension is a suspension of Nevirapine in
Inject the reference solution. The test is not valid unless the a suitable flavoured vehicle.
column efficiency determined from the nevirapine peak is not
less than 7500 theoretical plates and the tailing factor is not Nevirapine Oral Suspension contains not less than 90.0 per
more than 1.5 and the relative standard deviation for replicate cent and not more than 110.0 per cent of the stated amount of
injections is not more than 2 per cent. nevirapine, C15H14N4O.

Inject the test solution and continue the chromatography for Identification
at least five times the retention time of the principal peak.
A. Determine by thin-layer chromatography (2.4.17), coating
Determine the amount of related substances by the area
the plate with silica gel GF254.
normalisation method. Any individual impurity is not more
than 1.0 per cent and the sum of all impurities is not more than Mobile phase. A mixture of 40 volumes of 1-butanol,
2.0 per cent. 30 volumes of heptane, 30 volumes of acetone and 10 volumes
of strong ammonia solution.
Other tests. Comply with the tests stated under Tablets.
Test solution. Dilute the preparation under examination with
Assay. Determine by liquid chromatography (2.4.14).
methanol to obtain a solution containing 1 mg of Nevirapine
Test solution. Weigh and powder 20 tablets. Shake a quantity per ml.
of powder containing about 100 mg of Nevirapine with
Reference solution. A 0.1 per cent w/v solution of nevirapine
sufficient of the mobile phase to obtain a mixture containing
RS in a mixture of 75 volumes of methanol and 25 volumes of
0.05 per cent w/v of Nevirapine. Mix and filter through a
water.
membrane filter disc with an average pore diameter not greater
than 1.0 µm, rejecting the first few ml of the filtrate. Apply to the plate 10 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
Reference solution. A 0.05 per cent w/v solution of nevirapine The principal spot in the chromatogram obtained with the test
RS in the mobile phase. solution corresponds to that in the chromatogram obtained
Chromatographic system with the reference solution.
– a stainless steel column 25 cm x 4.6 mm, packed with
B. In the Assay, the principal peak in the chromatogram
octadecylsilane bonded to porous silica or ceramic
obtained with the test solution corresponds to the peak in the
microparticles (5 µm),
chromatogram obtained with the reference solution.
– mobile phase: a filtered and degassed mixture of
20 volumes of methanol, 20 volumes of acetonitrile Tests
and 60 volumes of a buffer prepared by dissolving
12.0 g of sodium dihydrogen phosphate in about 800 ml pH (2.4.24). 5.0 to 7.0.
of water, adjusting the pH to 3.0 with phosphoric acid Related substances. Determine by liquid chromatography
and diluting to 1000.0 ml with water, (2.4.14).

821
NICLOSAMIDE IP 2007

Test solution. To an accurately measured volume of the Reference solution. Weigh accurately about 50 mg of
preparation under examination containing about 25 mg of nevirapine RS, add about 20 ml of methanol, mix with the aid
Nevirapine add about 10 ml of methanol, mix with the aid of of ultrasound to dissolve and dilute to 100.0 ml with water.
ultrasound for 10 minutes, dilute to 50 ml with water, mix and Dilute 10.0 ml of this solution to 25.0 ml with water.
filter. Use the chromatographic system described under the test for
Reference solution. Weigh accurately about 25 mg of Related substances.
nevirapine RS, add about 10 ml of methanol, mix with the aid
Inject the reference solution. The test is not valid unless the
of ultrasound to dissolve and dilute to 50 ml with water.
relative standard deviation for replicate injections is not more
Chromatographic system than 2.0 per cent.
– a stainless steel column 25 cm x 4.6 mm, packed with
octylsilyl silica gel for chromatography (5 µm)(such as Inject separately the test solution and the reference solution
Hypersil C8), and measure the responses for the principal peak.
– mobile phase: filtered and degassed gradient mixtures Determine the weight per ml of the oral suspension (2.4.29)
of methanol and 0.1 M ammonium acetate, and calculate the content of C15H14N4O weight in volume.
– flow rate. 1 ml per minute,
Storage. Store protected from light.
– a linear gradient programme using the conditions given
below,
– spectrophotometer set at 270 nm,
– a 20 µl loop injector.
Niclosamide
Time 0.1 M ammonium acetate Methanol
(in min.) (per cent v/v) (per cent v/v) Anhydrous Niclosamide
0 95 5
5 95 5 Cl NO2
25 20 80 OH O
30 20 80 N
31 95 5 H
40 95 5
Inject the reference solution. The test is not valid unless the Cl
column efficiency determined from the nevirapine peak is not
less than 3000 theoretical plates and the tailing factor is not C13H8Cl2N2O4 Mol. Wt. 327.1
more than 2.0. Niclosamide is 2′,5-dichloro-4′-nitrosalicylanilide.
Inject separately the diluent (dilute 10 ml of methanol to 50 ml
Niclosamide contains not less than 98.0 per cent and not more
with water) and the test solution. Examine the diluent
than 101.0 per cent of C13H8Cl2N2O4, calculated on the dried
chromatogram for any extraneous peaks and ignore the
basis.
corresponding peaks observed in the chromatogram obtained
with the test solution. Ignore any peaks due to preservatives Description. A yellowish white to yellowish, fine crystals or
also. powder; odourless.
Any secondary peak observed in the chromatogram obtained
Identification
with the test solution should not be more than 1.0 per cent
and the sum of the areas of all the secondary peaks should Test A may be omitted if tests B and C are carried out. Tests B
not be more than 2.0 per cent when calculated by percentage and C may be omitted if test A is carried out.
area normalisation.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Other tests. Comply with the tests stated under Oral Liquids. Compare the spectrum with that obtained with niclosamide
Assay. Determine by liquid chromatography (2.4.14) RS or with the reference spectrum of niclosamide.
Test solution. Weigh accurately a quantity of the preparation B. Heat 50 mg with 5 ml of 1 M hydrochloric acid and 0.1 g of
under examination containing 25 mg of Nevirapine, add about zinc powder in a water-bath for 10 minutes, cool and filter. To
10 ml of methanol, mix with the aid of ultrasound for 10 minutes, the filtrate add 1 ml of a 0.5 per cent w/v solution of sodium
dilute to 50.0 ml with water, mix and filter. Further dilute 10.0 ml nitrite and allow to stand for 3 minutes. Add 2 ml of a 2 per
of the filtrate to 25.0 ml with water. cent w/v solution of ammonium sulphamate, shake, allow to

822
IP 2007 NICLOSAMIDE TABLETS

stand for 3 minutes and add 2 ml of a 0.5 per cent w/v solution Niclosamide Tablets
of N- (1-naphthyl) ethylenediamine dihydrochloride; a violet
colour is produced. Niclosamide Tablets contain not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of
C. Heat the substance under examination on a copper wire in niclosamide, C 13 H 8 Cl 2 N 2O 4 . The tablets may contain
a non-luminous flame; a green colour is imparted to the flame. sweetening and flavouring agents.
Tests Identification
Chlorides (2.3.12). To 2.0 g add a mixture of 40 ml of water and Heat a quantity of the powdered tablets containing 0.5 g of
1.2 ml of 5 M acetic acid, boil for 2 minutes, cool and filter; Niclosamide with 25 ml of hot ethanol (95 per cent), filter
10 ml of the filtrate diluted to 15 ml with water complies with while hot and evaporate the filtrate to dryness on a water-
the limit test for chlorides (500 ppm). bath. The residue complies with the following tests.
2-Chloro-4-nitroaniline. Not more than 100 ppm, determined A. Determine by infrared absorption spectrophotometry (2.4.6).
by the following method. Boil 0.25 g with 5 ml of methanol, Compare the spectrum with that obtained with niclosamide
cool, add 45 ml of 1 M hydrochloric acid, heat to boiling, RS or with the reference spectrum of niclosamide.
cool, filter and dilute the filtrate to 50.0 ml with 1 M
hydrochloric acid. To 10.0 ml of this solution add 0.5 ml of a B. Heat 50 mg with 5 ml of 1 M hydrochloric acid and 0.1 g of
0.5 per cent w/v solution of sodium nitrite and allow to stand zinc powder in a water-bath for 10 minutes, cool and filter. To
for 3 minutes. Add 1.0 ml of a 2 per cent w/v solution of the filtrate add 1 ml of a 0.5 per cent w/v solution of sodium
ammonium sulphamate, shake, allow to stand for 3 minutes nitrite and allow to stand for 3 minutes. Add 2 ml of a 2 per
and add 1.0 ml of a 0.5 per cent w/v solution of N- (1-naphthyl) cent w/v solution of ammonium sulphamate, shake, allow to
ethylenediamine dihydrochloride. Any pinkish violet colour stand for 3 minutes and add 2 ml of a 0.5 per cent w/v solution
produced is not more intense than that obtained in a solution of N- (1-naphthyl) ethylenediamine dihydrochloride; a violet
prepared at the same time and in the same manner using colour is produced.
10.0 ml of a solution prepared by diluting 2.0 ml of a
0.00050 per cent w/v solution of 2-chloro-4-nitroaniline in Tests
methanol to 20 ml with 1 M hydrochloric acid and beginning 2-Chloro-4-nitroaniline. Not more than 100 ppm, Boil a
at the words “add 0.5 ml of a 0.5 per cent w/v solution of quantity of the powdered tablets containing 0.1 g of
sodium nitrite.....”. Niclosamide with 20 ml of methanol and 20 ml of a solution in
5-Chlorosalicylic acid. Not more than 60 ppm, determined by methanol containing 10 µg of 2-chloro-4-nitroaniline, cool,
the following method. Boil 1.0 g with 15 ml of water for add 45 ml of 1 M hydrochloric acid, heat to boiling, cool, filter
2 minutes, cool, filter through a membrane filter (pore size and dilute the filtrate to 50.0 ml with 1 M hydrochloric acid.
0.45 µm), wash the filter and dilute the combined filtrate and To 10.0 ml of this solution add 0.5 ml of a 0.5 per cent w/v
washings to 20 ml with water (solution A). Dissolve 30 mg of solution of sodium nitrite and allow to stand for 3 minutes.
5-chlorosalicylic acid in 20 ml of methanol and add sufficient Add 1.0 ml of a 2 per cent w/v solution of ammonium
water to produce 100.0 ml. Dilute 1.0 ml of this solution to sulphamate, shake, allow to stand for 3 minutes and add
100.0 ml with water (solution B). To 10.0 ml of each of solutions 1.0 ml of a 0.5 per cent w/v solution of N- (1-naphthyl)
A and B add separately 0.1 ml of ferric chloride solution; any ethylenediamine dihydrochloride. Any pinkish violet colour
violet colour produced in solution A is not more intense than produced is not more intense than that obtained in a solution
that obtained in solution B. prepared at the same time and in the same manner using
10.0 ml of a solution prepared by diluting 2.0 ml of a 0.0005 per
Sulphated ash (2.3.18). Not more than 0.1 per cent.
cent w/v solution of 2-chloro-4-nitroaniline in methanol to
Loss on drying (2.4.19). Not more than 0.5 per cent, determined 20.0 ml with 1 M hydrochloric acid and beginning at the
on 1.0 g by drying in an oven at 105° for 4 hours. words “add 0.5 ml of a 0.5 per cent w/v solution of sodium
nitrite.....”.
Assay. Weigh accurately about 0.3 g, dissolve in 80 ml of a
mixture of equal volumes of acetone and methanol. Titrate 5-Chlorosalicylic acid. Boil a quantity of the powdered tablets
with 0.1 M tetrabutylammonium hydroxide, determining the containing 0.5 g of Niclosamide with 10 ml of water for
end-point potentiometrically (2.4.25). Carry out a blank titration. 2 minutes, cool, filter and to the filtrate add 0.2 ml of ferric
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to chloride solution; no red or violet colour is produced.
0.03271 g of C13H8Cl2N2O4. Disintegration. The test does not apply.
Storage. Store protected from light and moisture. Other Tests. Comply with the tests stated under Tablets.

823
NICLOTINAMIDE IP 2007

Assay. Weigh and powder 20 tablets. Weigh accurately a Mobile phase. A mixture of 48 volumes of chloroform,
quantity of the powdered tablets containing about 0.3 g of 45 volumes of ethanol and 4 volumes of water.
Niclosamide dissolved in 60 ml of dimethylformamide. Titrate Test solution. Dissolve 0.8 g of the substance under
with 0.1 M tetrabutylammonium hydroxide, determining the examination in 10 ml of ethanol (50 per cent).
end-point potentiometrically (2.4.25). Carry out a blank titration.
Reference solution. A 0.02 per cent w/v solution of the
1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to substance under examination in ethanol (50 per cent).
0.03271 g of C13H8Cl2N2O4.
Apply to the plate 5 µl of each solution. Allow the mobile
Storage. Store protected from light and moisture. phase to rise 10 cm. Dry the plate in air and examine in ultraviolet
Labelling. The label states that the tablets should be chewed light at 254 nm. Any secondary spot in the chromatogram
thoroughly before swallowing. obtained with the test solution is not more intense than the
spot in the chromatogram obtained with the reference solution.
Heavy metals (2.3.13). Dissolve 0.67 g in 10 ml of water, 7.5 ml
Nicotinamide of 1 M hydrochloric acid and sufficient water to produce
25 ml. The solution complies with the limit test for heavy metals,
Niacinamide Method A (30 ppm).
N Chlorides (2.3.12). 1.0 g complies with the limit test for
chlorides (250 ppm).
NH2
Sulphates (2.3.17). 1.2 g complies with the limit test for
O sulphates (125 ppm).
C6H6N2O Mol. Wt. 122.1 Sulphated ash (2.3.18). Not more than 0.1 per cent.
Nicotinamide is pyridine-3-carboxamide. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
on 1.0 g by drying over phosphorus pentoxide at a pressure
Nicotinamide contains not less than 99.0 per cent and not
of 1.5 to 2.7 kPa for 18 hours.
more than 101.0 per cent of C6H6N2O, calculated on the dried
basis. Assay. Weigh accurately about 0.25 g, dissolve in 20 ml of
anhydrous glacial acetic acid, heating slightly if necessary.
Description. Colourless crystals or a white, crystalline powder;
Add 5 ml of acetic anhydride. Titrate with 0.1 M perchloric
odour, faint and characteristic.
acid, using crystal violet solution as indicator. Carry out a
Identification blank titration.

Test A may be omitted if tests B, C and D are carried out. Tests 1 ml of 0.1 M perchloric acid is equivalent to 0.01221 g of
B, C and D may be omitted if test A is carried out. C6H6N2O.
A. Determine by infrared absorption spectrophotometry (2.4.6). Storage. Store protected from moisture.
Compare the spectrum with that obtained with nicotinamide
RS or with the reference spectrum of nicotinamide.
B. Heat about 5 mg in a dry tube; pyridine is evolved.
Nicotinamide Tablets
C. Boil 0.1 g with 1 ml of dilute sodium hydroxide solution; Niacinamide Tablets
ammonia is evolved. Nicotinamide Tablets contain not less than 92.5 per cent and
D. To 2 ml of a 0.1 per cent w/v solution add 6 ml of cyanogen not more than 107.5 per cent of the stated amount of
bromide solution and 1 ml of a 2.5 per cent w/v solution of nicotinamide, C6H6N2O.
aniline; a golden yellow colour develops.
Identification
Tests
Shake a quantity of the powdered tablets containing 0.2 g of
Appearance of solution. A 5.0 per cent w/v solution in carbon Nicotinamide with 50 ml of ethanol for 15 minutes, filter and
dioxide-free water is clear (2.4.1), and not more intensely evaporate the filtrate to dryness on a water-bath. The residue
coloured than reference solution BYS7 (2.4.1). complies with the following tests.
pH (2.4.24). 6.0 to 7.5, determined in a 5.0 per cent w/v solution. A. Determine by infrared absorption spectrophotometry (2.4.6).
Related substances. Determine by thin-layer chromatography Compare the spectrum with that obtained with nicotinamide
(2.4.17), coating the plate with silica gel GF254. RS or with the reference spectrum of nicotinamide.

824
IP 2007 NICLOTINIC ACID

B. Boil 0.1 g with 1 ml of dilute sodium hydroxide solution; Nicotinic Acid contains not less than 99.5 per cent and not
ammonia is evolved. more than 100.5 per cent of C6H5NO2, calculated on the dried
C. To 2 ml of a 0.1 per cent w/v solution add 6 ml of cyanogen basis.
bromide solution and 1 ml of a 2.5 per cent w/v solution of Description. A white or creamy-white, crystalline powder.
aniline; a golden yellow colour develops.
D. When examined in the range 230 nm to 360 nm (2.4.7), the
Identification
solution obtained in the Assay shows an absorption maximum Test A may be omitted if tests B, C and D are carried out. Tests
only at about 262 nm and two shoulders at about 258 nm and B, C and D may be omitted if test A is carried out.
269 nm.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Tests Compare the spectrum with that obtained with nicotinic acid
RS or with the reference spectrum of nicotinic acid.
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel GF254. B. Heat a small quantity with twice its weight of soda lime;
pyridine is evolved.
Mobile phase. A mixture of 48 volumes of chloroform,
45 volumes of ethanol and 4 volumes of water. C. Dissolve about 50 mg in 20 ml of water, neutralise to litmus
paper with 0.1 M sodium hydroxide, add 3 ml of copper
Test solution. Shake a quantity of the powdered tablets sulphate solution; a blue precipitate is gradually produced.
containing 0.1 g of Nicotinamide with 15 ml of ethanol for
15 minutes, filter, evaporate to dryness on a water-bath and D. To 2 ml of a 0.1 per cent w/v solution add 6 ml of cyanogen
dissolve the residue as completely as possible in 1 ml of bromide solution and 1 ml of a 2.5 per cent w/v solution of
ethanol. aniline; a golden yellow colour is produced.

Reference solution. Dilute 1 volume of the test solution to Tests


400 volumes with ethanol
Related substances. Determine by thin-layer chromatography
Apply to the plate 5 µl of each solution. Allow the mobile
(2.4.17), coating the plate with silica gel GF254.
phase to rise 10 cm. Dry the plate in air and examine in ultraviolet
light at 254 nm. Any secondary spot in the chromatogram Mobile phase. A mixture of 85 volumes of 1-propanol,
obtained with the test solution is not more intense than the 10 volumes of anhydrous formic acid and 5 volumes of water.
spot in the chromatogram obtained with the reference solution. Test solution. Dissolve 0.2 g of the substance under
Other Tests. Comply with the tests stated under Tablets. examination in 10 ml of water. Warm slightly, if necessary.
Assay. Weigh and powder 20 tablets. Weigh accurately a Reference solution. A 0.01 per cent w/v solution of the
quantity of the powder containing about 50 mg of substance under examination in water.
Nicotinamide, shake with 50 ml of ethanol (95 per cent) for 15 Apply to the plate 5 µl of each solution. After development,
minutes and dilute to 100.0 ml with ethanol (95 per cent). Mix, dry the plate at 105° for 10 minutes and examine in ultraviolet
filter, dilute 5.0 ml of the filtrate to 100.0 ml with ethanol (95 light at 254 nm. Any secondary spot in the chromatogram
per cent) and measure the absorbance of the resulting solution obtained with the test solution is not more intense than the
at the maximum at about 262 nm (2.4.7). Calculate the content spot in the chromatogram obtained with the reference solution.
of C6H6N2O taking 241 as the specific absorbance at 262 nm.
Heavy metals (2.3.13). Mix 1.0 g with 1.5 ml of dilute
Storage. Store protected from light and moisture. hydrochloric acid and sufficient water to produce 25 ml, heat
gently and cool to room temperature. The resulting solution
complies with the limit test for heavy metals, Method B
Nicotinic Acid (20 ppm).
Chlorides (2.3.12). 1.0 g complies with the limit test for
Niacin
chlorides (250 ppm).
N Sulphated ash (2.3.18). Not more than 0.1 per cent.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined
COOH on 1.0 g by drying in an oven at 105° for 1 hour.
C6H5NO2 Mol. Wt. 123.1
Assay. Weigh accurately about 0.25 g, dissolve in 50 ml of
Nicotinic Acid is pyridine-3-carboxylic acid. carbon dioxide-free water and titrate with 0.1 M sodium

825
NICLOTINIC TABLETS IP 2007

hydroxide using phenol red solution as indicator. Repeat the stand for 15 minutes, swirling occasionally, and then shake
operation without the substance under examination. The for 10 minutes. Filter through a plug of cotton and wash the
difference between the titrations represents the amount of filter with ethanol (95 per cent). Add 50 ml of carbon dioxide-
sodium hydroxide required. free water and titrate with 0.1 M sodium hydroxide using
1 ml of 0.1 M sodium hydroxide is equivalent to 0.01231 g of phenol red solution as indicator.
C6H5NO2. 1 ml of 0.1 M sodium hydroxide is equivalent to 0.01231 g of
C6H5NO2.
Storage. Store protected from light and moisture.
Storage. Store protected from light and moisture.

Nicotinic Tablets
Niacin Acid Tablets Nicoumalone
Nicotinic Acid Tablets contain not less than 92.5 per cent and Acenocoumarol
not more than 107.5 per cent of the stated amount of nicotinic
acid, C6H5NO2. O O NO2

Identification
A. Determine by thin-layer chromatography (2.4.17), coating OH CH3
the plate with silica gel GF254.
O
Mobile phase. A mixture of 48 volumes of chloroform,
45 volumes of ethanol (95 per cent) and 8 volumes of water. C19H15NO6 Mol. Wt. 353.3
Test solution. Shake a quantity of the powdered tablets Nicoumalone is (RS)-4-hydroxy-3-[1-(4-nitrophenyl)-3-
containing 50 mg of Nicotinic Acid with 50 ml of hot ethanol oxobutyl]coumarin.
(95 per cent), filter and allow the filtrate to cool. Nicoumalone contains not less than 98.5 per cent and not
Reference solution. A 0.1 per cent w/v solution of nicotinic more than 100.5 per cent of C19H15NO6, calculated on the dried
acid RS in ethanol (95 per cent). basis.
Apply to the plate 5 µl of each solution. After development, Description. A white to brownish-white powder; odourless or
dry the plate in air and examine in ultraviolet light at 254 nm. almost odourless.
The principal spot in the chromatogram obtained with the test
solution corresponds to that in the chromatogram obtained Identification
with the reference solution. A. Determine by infrared absorption spectrophotometry (2.4.6).
B. Triturate a quantity of the powdered tablets containing Compare the spectrum with that obtained with nicoumalone
50 mg of Nicotinic Acid with 10 ml of water and filter. To 2 ml RS or with the reference spectrum of nicoumalone.
of the filtrate add 6 ml of cyanogen bromide solution and 1 ml B. When examined in the range 230 nm to 360 nm (2.4.7), a
of a 2.5 per cent w/v solution of aniline; a golden yellow 0.001 per cent w/v solution in a mixture of 9 volumes of
precipitate is produced. methanol and 1 volume of 1 M hydrochloric acid shows
C. Shake a quantity of the powdered tablets containing 0.1 g absorption maxima at about 283 nm and 306 nm; absorbances
of Nicotinic Acid with ethanol (95 per cent), filter and at the maxima, about 0.65 and about 0.52, respectively.
evaporate the filtrate to dryness. Add to the residue 10 mg of C. Heat 50 mg with 2.5 ml of glacial acetic acid, 0.5 ml of
citric acid and 0.15 ml of acetic anhydride and heat on a hydrochloric acid and 0.2 g of zinc powder on a water-bath
water-bath; a reddish-violet colour is produced. for 5 minutes, cool and filter. To the filtrate add 0.05 ml of
Tests sodium nitrite solution and add the mixture to 10 ml of a 1 per
cent w/v solution of 2-naphthol containing 3 ml of 5 M sodium
Other Tests. Comply with the tests stated under Tablets. hydroxide; a bright red precipitate is produced.
Assay. Weigh and powder 20 tablets. Weigh accurately a Tests
quantity of the powder containing about 0.25 g of Nicotinic
Acid, add 40 ml of hot ethanol (95 per cent), previously Appearance of solution. A 2.0 per cent w/v solution in acetone
neutralised to phenolphthalein solution, and shake. Allow to is clear (2.4.1).

826
IP 2007 NICOUMALONE TABLETS

B. A 2.0 per cent w/v solution in 0.1 M sodium hydroxide is On the residue determine by infrared absorption
clear (2.4.1), and yellow. spectrophotometry (2.4.6). Compare the spectrum with that
Related substances. Determine by thin-layer chromatography obtained with nicoumalone RS or with the reference spectrum
(2.4.17), coating the plate with silica gel GF254. of nicoumalone.
B. When examined in the range 230 nm to 360 nm (2.4.7), the
Mobile phase. A mixture of 50 volumes of chloroform,
final solution obtained in the Assay shows absorption maxima
50 volumes of cyclohexane and 20 volumes of glacial acetic
at about 283 nm and 306 nm.
acid.
C. Heat 50 mg of the residue obtained in test A, with 2.5 ml of
Test solution. Dissolve 0.2 g of the substance under
glacial acetic acid, 0.5 ml of hydrochloric acid and 0.2 g of
examination in 10 ml of acetone.
zinc powder on a water-bath for 5 minutes, cool and filter. To
Reference solution. A 0.002 per cent w/v solution of the the filtrate add 0.05 ml of sodium nitrite solution and add the
substance under examination in acetone. mixture to 10 ml of a 1 per cent w/v solution of 2-naphthol
Apply to the plate 20 µl of each solution. After development, containing 3 ml of 5 M sodium hydroxide; a bright red
dry the plate in air and immediately examine in ultraviolet light precipitate is produced.
at 254 nm. Any secondary spot in the chromatogram obtained
with the test solution is not more intense than the spot in the Tests
chromatogram obtained with the reference solution. Related substances. Determine by thin-layer chromatography
Sulphated ash (2.3.18). Not more than 0.1 per cent. (2.4.17), coating the plate with silica gel GF254.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined Mobile phase. A mixture of 50 volumes of chloroform,
on 1.0 g by drying in an oven at 105°. 50 volumes of cyclohexane and 20 volumes of glacial acetic
acid.
Assay. Weigh accurately about 0.75 g, dissolve in 50 ml of
acetone and titrate with 0.1 M sodium hydroxide using Test solution. Shake a quantity of the powdered tablets
bromothymol blue solution as indicator. Repeat the operation containing 20 mg of Nicoumalone with 5 ml of acetone,
without the substance under examination. The difference centrifuge and use the supernatant liquid.
between the titrations represents the amount of sodium Reference solution. Dilute 1 volume of the test solution to
hydroxide required. 200 volumes with acetone.
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03533 g of Apply to the plate 20 µl of each solution. After development,
C19H15NO6. dry the plate in air and immediately examine in ultraviolet light
Storage. Store protected from light. at 254 nm. Any secondary spot in the chromatogram obtained
with the test solution is not more intense than the spot in the
chromatogram obtained with the reference solution.
Uniformity of content. Comply with the test stated under
Nicoumalone Tablets Tablets.
Acenocoumarol Tablets Finely crush one tablet, add 30 ml of methanol, stir the mixture
Nicoumalone Tablets contain not less than 90.0 per cent and for 30 minutes and filter through sintered glass, washing the
not more than 110.0 per cent of the stated amount of residue with three quantities, each of 15 ml, of methanol. To
nicoumalone, C19H15NO6. the combined filtrate and washings add 10 ml of 1 M
hydrochloric acid and sufficient methanol to produce
Identification 100.0 ml. If necessary, dilute further with a solvent prepared
by diluting 1 volume of 1 M hydrochloric acid to 10 volumes
A. Heat a quantity of the powdered tablets containing 50 mg with methanol to produce a solution containing about
of Nicoumalone with 30 ml of acetone under a reflux condenser 0.001 per cent w/v solution of Nicoumalone. Measure the
for 5 minutes, filter and wash the residue with two quantities, absorbance of the resulting solution at the maximum at about
each of 10 ml, of acetone. Evaporate the combined filtrate and 306 nm (2.4.7). Calculate the content of C19H15NO6 taking 521
washings to 5 ml, add water dropwise until the solution as the specific absorbance at 306 nm.
becomes turbid, heat on a water-bath until the solution is
Other Tests. Comply with the tests stated under Tablets.
clear and allow to stand. Filter, wash the crystals with a mixture
of equal volumes of acetone and water and dry at 100° at a Assay. Weigh and powder 20 tablets. Weigh accurately a
pressure of 2 kPa for 30 minutes. quantity of the powder containing about 10 mg of

827
NIFEDIPINE IP 2007

Nicoumalone, add 30 ml of methanol, stir the mixture for the peak due to nifedipine in the chromatogram obtained with
30 minutes and filter through sintered-glass, washing the the reference solution.
residue with three quantities, each of 15 ml, of methanol. To C. Determine by thin-layer chromatography (2.4.17), coating
the combined filtrate and washings add 10 ml of 1 M the plate with silica gel GF254.
hydrochloric acid and sufficient methanol to produce
100.0 ml. Dilute 5.0 ml of this solution to 50.0 ml with a solvent Mobile phase. A mixture of 60 volumes of cyclohexane and
prepared by diluting 1 volume of 1 M hydrochloric acid to 40 volumes of ethyl acetate.
10 volumes with methanol and measure the absorbance of the Test solution. Dissolve 0.1 g of the substance under
resulting solution at the maximum at about 306 nm (2.4.7). examination in 100 ml of methanol.
Calculate the content of C19H15NO6 taking 521 as the specific
absorbance at 306 nm. Reference solution. A 0.1 per cent w/v solution of nifedipine
RS in methanol.
Storage. Store protected from light and moisture.
Apply to the plate 5 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
The principal spot in the chromatogram obtained with the test
solution corresponds to that in the chromatogram obtained
Nifedipine with the reference solution.
D. To 25 mg add 10 ml of a mixture of 5 volumes of ethanol
H (95 per cent), 3.5 volumes of water and 1.5 volumes of
H3C N CH3 hydrochloric acid and dissolve with gentle heating. Add
0.5 g of granulated zinc and allow to stand for 5 minutes,
H3CO OCH3
swirling occasionally. Filter, add 5 ml of a 1 per cent w/v solution
O O of sodium nitrite to the filtrate and allow to stand for 2 minutes.
NO2 Add 2 ml of a 5 per cent w/v solution of ammonium sulphamate,
shake vigorously with care and add 2 ml of a 0.5 per cent w/v
solution of N-(1- naphthyl) ethylenediamine dihydrochloride;
an intense red colour develops which persists for more than
C17H18N2O6 Mol. Wt. 346.3
5 minutes.
Nifedipine is dimethyl 1,4-dihydro-2,6-dimethyl-4-(2-
nitrophenyl)pyridine-3,5-dicarboxylate. Tests
Nifedipine contains not less than 98.0 per cent and not more Related substances. Determine by liquid chromatography
than 102.0 per cent of C17H18N2O6, calculated on the dried (2.4.14)
basis.
Test solution. Dissolve 0.2 g of the substance under
Description. A yellow, crystalline powder; readily affected by examination in 20 ml of methanol and dilute to 50 ml with the
exposure to light. mobile phase.
NOTE — Nifedipine, when exposed to daylight and certain Reference solution (a). Dissolve an accurately weighed
wavelengths of artificial light, readily converts to a quantity of nifedipine RS in sufficient methanol to produce a
nitrosophenyl derivative. Exposure to ultraviolet light leads 1.0 per cent w/v solution and dilute quantitatively with the
to the formation of a nitrophenyl derivative. Perform the mobile phase to obtain a 0.4 per cent w/v solution.
tests and assay in the dark or under long-wavelength light
Reference solution (b). A 0.04 per cent w/v solution of
(greater than 420 nm). Use low-actinic glassware.
dimethyl-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-
Identification dicaboxylate RS (nitrophenylpyridine analogue) in methanol.
Reference solution (c). A 0.04 per cent w/v solution of
Test A may be omitted if tests B, C and D are carried out. Tests
dimethyl-2,6-dimethyl-4-(2-nitrosophenyl)pyridine-3,5-
B, C and D may be omitted if test A is carried out.
dicarboxylate RS (nitroso- phenylpyridine analogue) in
A. Determine by infrared absorption spectrophotometry (2.4.6). methanol.
Compare the spectrum with that obtained with nifedipine RS
Reference solution (d). Mix 1 volume of each of reference
or with the reference spectrum of nifedipine.
solutions (b) and (c) and 0.1 volume of the test solution, dilute
B. In the test for Related substances, the principal peak in the to 10 volumes with the mobile phase and then dilute 2 volumes
chromatogram obtained with the test solution corresponds to of the resulting solution to 10 volumes with the mobile phase.

828
IP 2007 NIFEDIPINE CAPSULES

Chromatographic system Nifedipine Capsules


– a stainless steel column 15 cm x 4.6 mm, packed with
octadecylsilane bonded to porous silica (5 µm), Nifedipine Capsules contain not less than 90.0 per cent and
– mobile phase: 55 volumes of water, 36 volumes of not more than 110.0 per cent of the stated amount of nifedipine,
methanol and 9 volumes of acetonitrile, C17H18N2O6.
– flow rate. 1 ml per minute, NOTE — Nifedipine, when exposed to daylight and certain
– spectrophotometer set at 235 nm, wavelengths of artificial light, readily converts to a
– a 20 µl loop injector. nitrosophenyl derivative. Exposure to ultraviolet light leads
Inject reference solution (d). The peaks appear in the order to the formation of a nitrophenyl derivative. Perform the
nitrophenylpyridine analogue, nitrosophenylpyridine tests and assay in the dark or under long-wavelength light
analogue and nifedipine, which has a retention time of about (greater than 420 nm). Use low-actinic glassware.
15.5 minutes. The test is not valid unless, in the chromatogram
obtained with reference solution (d), (a) the resolution factor
Identification
between the peaks due to the nitrophenylpyridine analogue Determine by thin-layer chromatography (2.4.17), coating the
and the nitrosophenylpyridine analogue is greater than 1.5, plate with silica gel GF254.
(b) the resolution between the peaks due to the
Mobile phase. A mixture of equal volumes of ethyl acetate
nitrosophenylpyridine analogue and nifedipine is greater than
and cyclohexane.
1.5, and (c) the height of the peak due to the nitrophenyl-
pyridine analogue is at least 20 per cent of the full-scale Test solution. Transfer a quantity of the contents of the capsules
deflection. containing 30 mg of Nifedipine into a centrifuge tube containing
0.1 M sodium hydroxide, add 25 ml of dichloromethane,
Inject the test solution and reference solutions (a) and (d) and stopper the tube and shake gently for 1 hour. Centrifuge for
record the chromatograms for twice the retention time of 10 minutes at 2000 to 2500 rpm. Remove the supernatant
nifedipine. In the chromatogram obtained with the test solution aqueous layer by aspiration with a syringe and transfer 5 ml of
no secondary peak other than any peaks corresponding to the clarified lower layer to a suitable vial.
the nitrophenylpyridine analogue and the nitrosophenylpyridine
analogue has an area greater than that of the peak due to Reference solution (a). A 0.12 per cent w/v solution of
nifedipine in the chromatogram obtained with reference solution nifedipine RS in dichloromethane.
(d) and the areas of any peaks corresponding to the Reference solution (b). A mixture of equal volumes of test
nitrophenylpyridine analogue and the nitrosophenylpyridine solution and reference solution (a).
analogue are not greater than the areas of the corresponding
Apply to the plate 500 µl of each solution as bands 20 mm by
peaks in the chromatogram obtained with reference solution
3 mm. After development, dry the plate in air until the solvent
(d). The total amount of related substances is not greater than
is not detectable and immediately examine in ultraviolet light
0.3 per cent. Ignore any peak with an area less than 10 per cent
at 254 nm. The principal band, appearing as a dark blue band,
of the area of the peak due to nifedipine in the chromatogram
in the chromatogram obtained with the test solution
obtained with reference solution (d).
corresponds to that in the chromatogram obtained with the
Heavy metals (2.3.13). 2.0 g complies with limit test for heavy reference solution. Spray with a solution prepared in the
metals, Method B (10 ppm). following manner. Dissolve 3 g of bismuth subnitrate and 30 g
Sulphated ash (2.3.18). Not more than 0.1 per cent. of potassium iodide in 10 ml of 3 M hydrochloric acid and
dilute with water to 100 ml; dilute 10 ml to 100 ml with 0.3 M
Loss on drying (2.4.19). Not more than 0.5 per cent, determined hydrochloric acid. In the chromatogram obtained with test
on 1.0 g by drying in an oven at 105° for 2 hours. solution the principal band, appearing as a compact light
Assay. Weigh accurately about 0.13 g, dissolve in a mixture of orange band against a yellow background, corresponds to
25 ml of 2-methyl-2-propanol and 25 ml of 1 M perchloric that in the chromatogram obtained with reference solution (a).
acid and titrate with 0.1 M ceric ammonium sulphate, using The band obtained with reference solution (b) appears as a
0.1 ml of ferroin solution as indicator until the pink colour is single band under both visualisation procedures.
discharged, titrating slowly towards the end-point. Carry out Tests
a blank titration.
Uniformity of content. Comply with the test stated under
1 ml of 0.1 M ceric ammonium sulphate is equivalent to Capsules.
0.01732 g of C17H18N2O6.
Transfer the contents of a capsule quantitatively to a 200-ml
Storage. Store protected from light. volumetric flask with the aid of methanol, dilute to volume

829
NIFEDIPINE SUSTAINED-RELEASE TABLETS IP 2007

with methanol and mix. Complete the Assay beginning at the with the reference solution. Spray with a solution prepared in
words “Measure the absorbance....” and calculate the content the following manner. Dissolve 3 g of bismuth subnitrate and
of C17H18N2O6 in the capsule. 30 g of potassium iodide in 10 ml of 3 M hydrochloric acid
Other Tests. Comply with the tests stated under Capsules. and dilute to 100 ml with water; dilute 10 ml of this solution to
100 ml with 0.3 M hydrochloric acid. In the chromatogram
Assay. Transfer the contents of 5 capsules containing about obtained with the test solution the principal band, appearing
50 mg of Nifedipine quantitatively to a 200-ml volumetric flask as a compact light orange band against a yellow background,
with the aid of small quantities of methanol. Dilute to volume corresponds to that in the chromatogram obtained with
with methanol and mix. To 20.0 ml add sufficient methanol to reference solution.
produce 100.0 ml and mix. Measure the absorbance of the
resulting solution at the maximum at about 350 nm (2.4.7). Tests
Calculate the content of C17H18N2O6 in the capsules from the
absorbance obtained by repeating the operation with a Dissolution (2.5.2)
0.005 per cent w/v solution of nifedipine RS in methanol. A. Apparatus No. 1
Storage. Store protected from light. Medium. 900 ml of 0.1 M hydrochloric acid
Speed and time. 150 rpm and 120 minutes.
Withdraw a suitable volume of the medium and filter. Measure
the absorbance of the filtrate, suitably diluted with the
Nifedipine Sustained-release Tablets dissolution medium, if necessary, at the maximum at about 340
nm (2.4.7).
Nifedipine Sustained-release Tablets contain not less than
90.0 per cent and not more than 110.0 per cent of the stated Calculate the content of C17H18N2O6 in the medium from the
amount of nifedipine, C17H18N2O6. absorbance obtained from a solution of known concentration
of nifedipine RS in the same medium.
NOTE - Nifedipine, when exposed to daylight and certain
wavelengths of artificial light, readily converts to a D. Not less than 25 per cent and not more than 45 per cent of
nitrosophenyl derivative. Exposure to ultraviolet light leads the stated amount of C17H18N2O6.
to the formation of a nitrophenyl derivative. Perform the
B. Apparatus No. 1
tests and the assay in the dark or under long-wavelength
light (greater than 420 nm). Use low-actinic glassware. Medium. 900 ml of phosphate buffer pH 6.8
Speed and time. 150 rpm and 6 hours.
Identification
Withdraw a suitable volume of the medium and filter. Measure
Determine by thin-layer chromatography (2.4.17), coating the the absorbance of the filtrate, suitably diluted with the
plate with silica gel GF254. dissolution medium, if necessary, at the maximum at about 340
Mobile phase. A mixture of equal volumes of ethyl acetate nm (2.4.7).
and cyclohexane. Calculate the content of C17H18N2O6 in the medium from the
Test solution. Transfer a quantity of the contents of the capsules absorbance obtained from a solution of known concentration
containing 30 mg of Nifedipine into a centrifuge tube containing of nifedipine RS in the same medium.
0.1 M sodium hydroxide, add 25 ml of dichloromethane,
D. Not less than 60 per cent of the stated amount of
stopper the tube and shake gently for 1 hour. Centrifuge for 10
C17H18N2O6.
minutes at 2000 rpm to 2500 rpm. Remove the supernatant
aqueous layer by aspiration with a syringe and use 5 ml of the Other tests. Comply with the tests stated under Tablets.
clarified lower layer.
Assay. Weigh and powder 20 tablets. Weigh accurately a
Reference solution. A 0.12 per cent w/v solution of nifedipine quantity of the powder containing about 25 mg of Nifedipine,
RS in dichloromethane. disperse in methanol, shake and dilute to 100.0 ml with
Apply to the plate 500 µl of each solution as bands 20 mm by methanol, filter. Dilute 20.0 ml of the filtrate to 100.0 ml with
3 mm. After development, dry the plate in air until the odour of methanol. Measure the absorbance of the resulting solution
the solvent is not detectable and immediately examine in at the maximum at about 350 nm (2.4.7). Calculate the content
ultraviolet light at 254 nm. The principal band, appearing as a of C17H18N2O6 from the absorbance obtained with a 0.005 per
dark blue band, in the chromatogram obtained with the test cent w/v solution of nifedipine RS in methanol.
solution corresponds to that in the chromatogram obtained Storage. Store protected from light and moisture.

830
IP 2007 NIKETHAMIDE

Nifedipine Tablets Assay beginning at the words “Measure the absorbance....”


and calculate the content of C17H18N2O6 in the tablet.
Nifedipine Tablets contain not less than 90.0 per cent and not
more than 110.0 per cent of the stated amount of nifedipine, Other Tests. Comply with the tests stated under Tablets.
C17H18N2O6. The tablets may be coated. Assay. Weigh and powder 20 tablets. Weigh accurately a
NOTE — Nifedipine, when exposed to daylight and certain quantity of the powder containing 50 mg of Nifedipine into a
wavelengths of artificial light, readily converts to a 200-ml volumetric flask. Dissolve with the aid of 50 ml of
nitrosophenyl derivative. Exposure to ultraviolet light leads methanol. Dilute to volume with methanol, mix and filter. Dilute
to the formation of a nitrophenyl derivative. Perform the 20 ml of the filtrate to 100 ml with methanol and mix. Measure
tests and assay in the dark or under long-wavelength light the absorbance of the resulting solution at the maximum at
(greater than 420 nm). Use low-actinic glassware. about 350 nm (2.4.7). Calculate the content of C17H18N2O6 from
the absorbance obtained by repeating the operation with a
Identification 0.005 per cent w/v solution of nifedipine RS in methanol.
Determine by thin-layer chromatography (2.4.17), coating the Storage. Store protected from light and moisture.
plate with silica gel GF254.
Mobile phase. A mixture of equal volumes of ethyl acetate
and cyclohexane.
Test solution. Transfer a quantity of the powdered tablets
Nikethamide
containing 30 mg of Nifedipine to a centrifuge tube containing
20 ml of 0.1 M sodium hydroxide, add 25 ml of dichloromethane, N CH3
stopper the tube and shake gently for 1 hour. Centrifuge for 10
minutes at 2000 to 2500 rpm. Remove the supernatant aqueous N CH3
layer by aspiration with a syringe and transfer 5.0 ml of the
clarified lower layer to a suitable vial. O

Reference solution (a). A 0.12 per cent w/v solution of


C10H14N2O Mol. Wt. 178.2
nifedipine RS in dichloromethane.
Nikethamide is N,N-diethylpyridine-3-carboxamide.
Reference solution (b). A mixture of equal volumes of the test
solution and reference solution (a). Nikethamide contains not less than 99.0 per cent and not more
than 101.0 per cent of C10H14N2O, calculated on the anhydrous
Apply to the plate 500 µl of each solution as bands 20 mm by
basis.
3 mm. After development, dry the plate in air until the solvent
is not detectable and immediately examine in ultraviolet light Description. A colourless or slightly yellowish, oily liquid or
at 254 nm. The principal band, appearing as a dark blue band, crystalline mass; odour, slight and characteristic.
in the chromatogram obtained with the test solution
corresponds to that in the chromatogram obtained with Identification
reference solution (a). Spray with a solution prepared in the
Test A may be omitted if tests B, C and D are carried out. Tests
following manner. Dissolve 3 g of bismuth subnitrate and 30 g
C and D may be omitted if tests A and B are carried out.
of potassium iodide in 10 ml of 3 M hydrochloric acid and
dilute with water to 100 ml; dilute 10 ml to 100 ml with 0.3 M A. Determine by infrared absorption spectrophotometry (2.4.6).
hydrochloric acid. In the chromatogram obtained with the Compare the spectrum with that obtained with nikethamide
test solution the principal band, appearing as a compact light RS or with the reference spectrum of nikethamide.
orange band against a yellow background, corresponds to
B. When examined in the range 230 nm to 360 nm (2.4.7), a
that in the chromatogram obtained with reference solution (a).
0.002 per cent w/v solution shows an absorption maximum
The band obtained with reference solution (b) appears as a
only at about 263 nm; absorbance at about 263 nm, about 0.57.
single band under both visualisation procedures.
C. Heat 0.1 g with 1 ml of 2 M sodium hydroxide; diethylamine,
Tests recognisable by its odour, is evolved progressively; the fumes
Uniformity of content. Comply with the test stated under turn red litmus paper blue.
Tablets D. To 2 ml of a 0.1 per cent w/v solution add 2 ml of cyanogen
Shake one tablet with methanol in a 200-ml volumetric flask, bromide solution and 3 ml of a 2.5 per cent w/v solution of
dilute to volume with methanol, mix and filter. Complete the aniline and mix; a yellow colour is produced.

831
NIKETHAMIDE INJETION IP 2007

Tests Identification
Appearance of solution. The substance, in liquid form or A. Make 1 ml alkaline with 5 M sodium hydroxide, extract with
liquefied by gentle heating, is clear (2.4.1), and not more 5 ml of dichloromethane and evaporate the solvent.
intensely coloured than reference solution YS5 (2.4.1).
On the residue determine by infrared absorption
pH (2.4.24). 6.0 to 7.8, determined in a 25.0 per cent w/v solution. spectrophotometry (2.4.6). Compare the spectrum with that
Congealing temperature (2.4.10). 23° to 24.5°. obtained with nikethamide RS or with the reference spectrum
of nikethamide.
Refractive index (2.4.27). 1.522 to1.526.
B. Gives a voluminous precipitate with alkaline potassium
Related substances. Determine by thin-layer chromatography mercuri-iodide solution and a greyish-brown flocculent
(2.4.17), coating the plate with silica gel GF254. precipitate with tannic acid solution. Gives no precipitate
Mobile phase. A mixture of 75 volumes of chloroform and with iodine solution or with potassium mercuri-iodide
25 volumes of 1-propanol. solution.
Test solution. Dissolve 0.4 g of the substance under C. Heat 1 ml with 0.2 g of sodium hydroxide; diethylamine,
examination in 10 ml of methanol. recognisable by its odour, is evolved.
Reference solution (a). A 0.04 per cent w/v solution of
ethylnicotinamide RS in methanol. Tests
Reference solution (b). A 0.004 per cent w/v solution of pH (2.4.24). 6.0 to 8.0.
ethylnicotinamide RS in methanol.
Related substances. Determine by thin-layer chromatography
Apply to the plate 10 µl of each solution. After development, (2.4.17), coating the plate with silica gel GF254.
dry the plate in air and examine in ultraviolet light at 254 nm.
Mobile phase. A mixture of 75 volumes of chloroform and
Any spot corresponding to ethylnicotinamide in the
25 volumes of 1-propanol.
chromatogram obtained with the test solution is not more
intense than the spot in the chromatogram obtained with Test solution. Dilute 1 ml of the injection to 5 ml with methanol.
reference solution (a) and any other secondary spot is not
Reference solution (a). A 0.05 per cent w/v solution of
more intense than the spot in the chromatogram obtained
ethylnicotinamide RS in methanol.
with reference solution (b).
Heavy metals (2.3.13). 2.0 g complies with limit test for heavy Reference solution (b). A 0.005 per cent w/v solution of
metals, Method B (10 ppm). ethylnicotinamide RS in methanol.

Sulphated ash (2.3.18). Not more than 0.1 per cent. Apply to the plate 10 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
Water (2.3.43). Not more than 0.3 per cent, determined on Any spot corresponding to ethylnicotinamide in the
2.0 g. chromatogram obtained with the test solution is not more
Assay. Weigh accurately about 0.15 g, dissolve in 20 ml of intense than the spot in the chromatogram obtained with
anhydrous glacial acetic acid and 5 ml of acetic anhydride. reference solution (a) and any other secondary spot is not
Titrate with 0.1 M perchloric acid, determining the end-point more intense than the spot in the chromatogram obtained
potentiometrically (2.4.25). Carry out a blank titration. with reference solution (b).
1 ml of 0.1 M perchloric acid is equivalent to 0.01782 g of Other tests. Complies with the tests stated under Parenteral
C10H14N2O. Preparations (Injections).
Storge. Store protected from light. Assay. Dilute 5.0 ml to 500.0 ml with water. To 5.0 ml of the
solution add 5 ml of 1 M hydrochloric acid and sufficient
water to produce 500.0 ml. Measure the absorbance of the
Nikethamide Injection resulting solution at the maximum at about 263 nm (2.4.7).
Calculate the content of C10H14N2O taking 282 as the specific
Nikethamide Injection is a sterile solution containing 25 per absorbance at 263 nm.
cent w/v solution of Nikethamide in Water for Injections.
Storage. Store protected from light, in single dose containers.
Nikethamide Injection contains not less than 24.0 per cent
and not more than 26.0 per cent w/v solution of nikethamide,
C10H14N2O.

832
IP 2007 NITRAZEPAM TABLETS

Nitrazepam Test solution. Dissolve 0.2 g of the substance under


examination in 10 ml of acetone.
O Reference solution. A 0.002 per cent w/v solution of the
HN substance under examination in acetone.
Apply to the plate 10 µl of each solution. Allow the mobile
N phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
O2N light at 254 nm. Any secondary spot in the chromatogram
obtained with the test solution is not more intense than the
spot in the chromatogram obtained with the reference solution.
Heavy metals (2.3.13). 1.0 g complies with the limit test for
C15H11N3O3 Mol. Wt. 281.3 heavy metals, Method B (20 ppm).
Nitrazepam is 1,3-dihydro-7-nitro-5-phenyl-2H-1,4- Sulphated ash (2.3.18). Not more than 0.1 per cent.
benzodiazepin-2-one. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Nitrazepam contains not less than 99.0 per cent and not more on 1.0 g by drying in an oven at 105° for 4 hours.
than 101.0 per cent C15H11N3O3, calculated on the dried basis. Assay. Weigh accurately about 0.25 g, dissolve in 50 ml of
Description. A yellow, crystalline powder; odourless or almost acetic anhydride. Titrate with 0.1 M perchloric acid,
odourless. determining the end-point potentiometrically (2.4.25). Carry
out a blank titration.
Identification
1 ml of 0.1 M perchloric acid is equivalent to 0.02813 g of
Test A may be omitted if tests B, C and D are carried out. Tests C15H11N3O3.
B, C and D may be omitted if test A is carried out. Storage. Store protected from light and moisture.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with nitrazepam RS
or with the reference spectrum of nitrazepam. Nitrazepam Tablets
B. Carry out the following procedure in subdued light.
Nitrazepam Tablets contain not less than 90.0 per cent and not
When examined in the range 230 nm to 360 nm (2.4.7), a freshly more than 110.0 per cent of the stated amount of nitrazepam,
prepared 0.0005 per cent w/v solution in a 0.5 per cent w/v C15H11N3O3.
solution of sulphuric acid in methanol shows an absorption
maximum only at about 280 nm; absorbance at about 280 nm, Identification
about 0.45. Carry out the following procedure in subdued light.
C. Dissolve 10 mg in 1 ml of methanol, warming if necessary, A. When examined in the range 230 nm to 360 nm (2.4.7), the
and add 0.05 ml of 2 M sodium hydroxide; an intense yellow final solution obtained in the Assay shows an absorption
colour is produced. maximum only at about 280 nm.
D. Dissolve 20 mg in a mixture of 5 ml of hydrochloric acid B. Determine by thin-layer chromatography (2.4.17), coating
and 10 ml of water, boil for 5 minutes, cool and add 2 ml of a the plate with silica gel G.
0.1 per cent w/v solution of sodium nitrite. Allow to stand for
1 minute, add 1 ml of a 0.5 per cent w/v solution of sulphamic Mobile phase. A mixture of 100 volumes of chloroform and
acid, mix, allow to stand for 1 minute, add 1 ml of a 0.1 per cent 10 volumes of methanol.
w/v solution of N-(1-naphthyl)ethylenediamine Test solution. Shake a quantity of the powdered tablets with
dihydrochloride; a red colour is produced. sufficient methanol to produce a solution containing 5 mg of
Nitrazepam per ml, allow to settle and decant the supernatant
Tests liquid.
Related substances and decomposition products. Determine Reference solution. A 0.5 per cent w/v solution of nitrazepam
by thin-layer chromatography (2.4.17), coating the plate with RS in methanol.
silica gel GF254.
Apply to the plate 2 µl of each solution. After development,
Mobile phase. A mixture of 85 volumes of nitromethane and dry the plate in air, spray it with ethanolic sulphuric acid
15 volumes of ethyl acetate. (10 per cent v/v), heat at 105° for 10 minutes and examine in

833
NITROFURANTOIN IP 2007

ultraviolet light at 365 nm. The principal spot in the light. Add 70 ml of a 0.5 per cent v/v solution of hydrochloric
chromatogram obtained with the test solution corresponds to acid in methanol, shake for 15 minutes protected from light,
that in the chromatogram obtained with the reference solution. add sufficient of the hydrochloric acid solution to produce
C. To a quantity of the powdered tablets containing 5 mg of 100.0 ml and filter. Dilute 10.0 ml of the filtrate to 100.0 ml with
Nitrazepam add 5 ml of hydrochloric acid and 10 ml of water, the same solvent and measure the absorbance of the resulting
heat on a water-bath for 15 minutes, filter and cool. To the solution immediately at the maximum at about 280 nm (2.4.7).
clear filtrate add 1 ml of a 0.1 per cent w/v solution of sodium Calculate the content of C15H11N3O3 taking 910 as the specific
nitrite, allow to stand for 3 minutes and add 1 ml of a 0.5 per absorbance at 280 nm.
cent w/v solution of sulphamic acid. Allow to stand for Storage. Store protected from light and moisture.
3 minutes and add 1 ml of a 0.1 per cent w/v solution of N-(1-
naphthyl) ethylenediamine dihydrochloride; a red colour is
produced.
Nitrofurantoin
Tests
O
Related substances and decomposition products. Determine H
by thin-layer chromatography (2.4.17), coating the plate with O 2N O C N N NH
silica gel GF254.
Mobile phase. A mixture of 40 volumes of nitromethane, O
40 volumes of toluene and 20 volumes of chloroform. C8H6N4O5 Mol. Wt. 238.2 (anhydrous)
Test solution. Shake a quantity of the powdered tablets C8H6N4O5,H2O Mol. Wt. 256.2 (hydrous)
containing 40 mg of Nitrazepam with 25 ml of chloroform,
Nitrofurantoin is 1-(5-nitrofurfurylideneamino)imidazolidine-
filter, carefully evaporate the filtrate to dryness and dissolve
2,4-dione. It is anhydrous or contains one molecule of water
the residue in 2 ml of chloroform.
of hydration.
Reference solution. Dilute 1 volume of the test solution to
200 volumes with chloroform. Nitrofurantoin contains not less than 98.0 per cent and not
more than 102.0 per cent of C8H6N4O5, calculated on the dried
Apply to the plate 5 µl of each solution. Allow the mobile basis.
phase to rise 12 cm. Dry the plate in air and examine in ultraviolet
light at 254 nm. Any secondary spot in the chromatogram Description. Lemon yellow crystals or a crystalline powder;
obtained with the test solution is not more intense than the odourless or almost odourless.
spot in the chromatogram obtained with the reference solution.
Identification
Uniformity of content. Comply with the test stated under
Tablets. Carry out the following test in subdued light.

NOTE — Carry out the following procedure in subdued light. A. When examined in the range 230 nm to 400 nm (2.4.7), the
final solution obtained in the Assay shows absorption maxima
Powder 1 tablet, add 5 ml of water, mix and allow to stand for
at about 266 nm and 367 nm; the ratio of the absorbance at the
15 minutes protected from light. Add 70 ml of a 0.5 per cent
maximum at about 367 nm to that at the maximum at about
v/v solution of hydrochloric acid in methanol, shake for
266 nm is 1.36 to 1.42.
15 minutes protected from light, add sufficient of the
hydrochloric acid solution to produce 100.0 ml and filter. B. To 1 ml of a 0.1 per cent w/v solution in dimethylformamide
Dilute 10.0 ml of the filtrate with sufficient of the hydrochloric add 0.1 ml of 0.5 M ethanolic potassium hydroxide; a brown
acid solution to produce a solution containing 0.0005 per colour develops.
cent w/v solution of Nitrazepam. Measure the absorbance of
the resulting solution immediately at the maximum at about
Tests
280 nm (2.4.7). Calculate the content of C15H11N3O3 in the tablet Related substances. Determine by thin-layer chromatography
taking 910 as the specific absorbance at 280 nm. (2.4.17), coating the plate with silica gel HF254.
Other Tests. Comply with the tests stated under Tablets. Mobile phase. A mixture of 90 volumes of nitromethane and
Assay. Carry out the following procedure in subdued light. 10 volumes of methanol.
Weigh and powder 20 tablets. Weigh accurately a quantity of Test solution. Dissolve 0.25 g of the substance under
the powder containing about 5 mg of Nitrazepam, add 5 ml of examination in minimum volume of dimethylformamide and
water, mix and allow to stand for 15 minutes protected from dilute to 10 ml with acetone.

834
IP 2007 NITROFURAZONE

Reference solution. Dilute 1 volume of the test solution to Reference solution. Dilute 1 volume of the test solution to
100 volumes with acetone. 100 volumes with acetone.
Apply to the plate 10 µl of each solution. After development, Apply to the plate 10 µl of each solution. After development,
dry the plate in air, heat at 105° for 5 minutes and examine in dry the plate in air, heat at 105° for 5 minutes and examine in
ultraviolet light at 254 nm. Spray with phenylhydrazine ultraviolet light at 254 nm. Spray with phenylhydrazine
hydrochloride solution and heat the plate at 105° for further hydrochloride solution and heat the plate at 105° for further
10 minutes. Any secondary spot in the chromatogram obtained 10 minutes. Any secondary spot in the chromatogram obtained
with the test solution is not more intense than the spot in the with the test solution, is not more intense than the spot in the
chromatogram obtained with the reference solution by both chromatogram obtained with the reference solution by both
methods of visualisation. methods of visualisation.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Other Tests. Comply with the tests stated under Tablets.
Loss on drying (2.4.19). 5.0 to 7.1 per cent (hydrous form) and Assay. Carry out the following procedure in subdued light.
not more than 1.0 per cent (anhydrous form), determined on
1.0 g by drying in an oven at 105°. Weigh and powder 20 tablets. Weigh accurately a quantity of
the powder containing about 0.15 g of Nitrofurantoin, add
Assay. Carry out the following procedure in subdued light. 50.0 ml of dimethylformamide, shake for 5 minutes, add
Weigh accurately about 75 mg and dissolve in 25.0 ml of sufficient water to produce 1000.0 ml and mix. Dilute 5.0 ml to
dimethylformamide, add sufficient water to produce 500.0 ml 100.0 ml with a solution containing 1.8 per cent w/v solution
and mix. Dilute 5.0 ml to 100.0 ml with a solution containing of sodium acetate and 0.14 per cent v/v of glacial acetic
1.8 per cent w/v solution of sodium acetate and 0.14 per cent acid. Measure the absorbance of the resulting solution at the
v/v of glacial acetic acid. Measure the absorbance of the maximum at about 367 nm (2.4.7), using as the blank the sodium
resulting solution at the maximum at about 367 nm (2.4.7), acetate-acetic acid solution. Calculate the content of C8H6N4O5
using as the blank the sodium acetate-acetic acid solution. taking 765 as the specific absorbance at 367 nm.
Calculate the content of C8H6N4O5 taking 765 as the specific
Storage. Store protected from light and moisture.
absorbance at 367 nm.
Storage. Store protected from light and moisture.
Labelling. The label states whether the material is anhydrous
or hydrous.
Nitrofurazone
Nitofural

Nitrofurantoin Tablets H
H N NH2
Nitrofurantoin Tablets contain not less than 90.0 per cent and O2N O C N
not more than 110.0 per cent of the stated amount of O
nitrofurantoin, C8H6N4O5.
C6H6N4O4 Mol. Wt. 198.1
Identification
Nitrofurazone is 5-nitro-2-furaldehyde semicarbazone.
Carry out the following procedure in subdued light.
Nitrofurazone contains not less than 97.0 per cent and not
A. When examined in the range 230 nm to 400 nm (2.4.7), the more than 103.0 per cent of C6H6N4O4, calculated on the dried
final solution obtained in the Assay shows absorption maxima basis.
at about 266 nm and 367 nm.
Description. A yellow to brownish-yellow, crystalline powder;
Tests odourless or almost odourless.
Related substances. Determine by thin-layer chromatography
Identification
(2.4.17), coating the plate with silica gel HF254.
Mobile phase. A mixture of 90 volumes of nitromethane and A. Determine by infrared absorption spectrophotometry (2.4.6).
10 volumes of methanol. Compare the spectrum with that obtained with nitrofurazone
RS or with the reference spectrum of nitrofurazone.
Test solution. Shake a quantity of the powdered tablets
containing 0.1 g of Nitrofurantoin with 10 ml of a mixture of B. Dissolve 1 mg in 1 ml of dimethylformamide and add 0.05 ml
9 volumes of acetone and 1 volume of dimethylformamide of 1 M ethanolic potassium hydroxide; a ruby red colour is
and filter. produced.

835
NITROUS OXIDE IP 2007

Tests less than 6 hours before carrying out the tests Keep the
cylinder in the vertical position with the outlet valve
pH (2.4.24). 5.0 to 7.0, determined in the filtrate obtained by uppermost and deliver the gas at a rate of 4 litres per hour,
shaking 1.0 g with 100 ml of carbon dioxide-free water and unless otherwise directed. The test for Carbon monoxide
filtering. should be carried out on the first portion of gas drawn from
Related substances. Carry out the following procedure in the cylinder and that for Nitric oxide and nitrogen dioxide
subdued light. immediately thereafter.
Determine by thin-layer chromatography (2.4.17), coating the Description. A colourless gas; odourless.
plate with silica gel G.
Mobile phase. A mixture of 95 volumes of toluene and Identification
5 volumes of dioxan. A. A glowing splinter of wood bursts into flame on contact
Test solution. Dissolve 0.1 g of the substance under with the gas.
examination in 10 ml of a mixture of equal volumes of acetone B. Shake with alkaline pyrogallol solution; the gas being
and dimethylformamide. examined is not absorbed and the solution does not become
Reference solution (a). A 0.002 per cent w/v solution of brown.
5-nitrofurfurylidene azine RS in a mixture of equal volumes of
acetone and dimethylformamide. Tests
Reference solution (b). A 0.01 per cent w/v solution of Acidity or alkalinity. Use hermetically-closed, flat-bottomed,
nitrofurfural diacetate RS in a mixture of equal volumes of glass cylinders with dimensions such that 50 ml of liquid
acetone and dimethylformamide. reaches a height of 12 to 14 cm, fitted with an outlet tube and
Apply to the plate 10 µl of each solution. After development, with an inlet tube with an orifice of 1 mm in internal diameter
dry the plate in air, heat it at 105° for 5 minutes and spray with reaching to within 2 mm of the bottom of the cylinder. For
phenylhydrazine hydrochloride solution. In the solution (1) pass 2.0 litres of the gas under examination through
chromatogram obtained with the test solution any spots a mixture of 0.1 ml of 0.01 M hydrochloric acid and 50 ml of
corresponding to 5-nitrofurfurylidene azine and nitrofurfural carbon dioxide-free water. For solution (2) use 50 ml of carbon
diacetate are not more intense than the spots in the dioxide-free water. For solution (3) add 0.2 ml of 0.01 M
chromatograms obtained with reference solutions (a) and (b) hydrochloric acid to 50 ml of carbon dioxide-free water. To
respectively. each solution add 0.1 ml of a 0.02 per cent w/v solution of
methyl red in ethanol (70 per cent). The intensity of the colour
Sulphated ash (2.3.18). Not more than 0.1 per cent.
of solution (1) is between those of solutions (2) and (3).
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Arsine and phosphine. Through a mercuric chloride paper
on 1.0 g by drying in an oven at 105°.
attached to a glass tube as in the limit test for arsenic (2.3.10),
Assay. Carry out the following procedure in subdued light. pass 2.0 litres of the gas; no visible stain is produced.
Weigh accurately about 60 mg, add 20.0 ml of Carbon dioxide. Not more than 300 ppm v/v determined by the
dimethylformamide, swirl to dissolve and add sufficient water following method. Use the apparatus described in the test for
to produce 500.0 ml. Dilute 5.0 ml of the solution to 100.0 ml Acidity or alkalinity. Pass 1.0 litre through 50 ml of clear barium
with water and mix. Measure the absorbance of the resulting hydroxide solution. Any turbidity produced in the resulting
solution at the maximum at about 375 nm (2.4.7). Calculate the solution is not more than that obtained in a reference solution
content of C6H6N4O4 taking 822 as the specific absorbance at prepared at the same time by adding 1 ml of a 0.11 per cent
375 nm. w/v solution of sodium bicarbonate in carbon dioxide-free
Storage. Store protected from light and moisture. water to 50 ml of barium hydroxide solution.
Carbon monoxide. Not more than 10 ppm v/v, determined by
the following method. Connect in series a U-tube containing
Nitrous Oxide silica gel impregnated with chromium trioxide, a drechsel
N2O Mol. Wt. 44.0 bottle containing 100 ml of a 40 per cent w/v solution of
potassium hydroxide, a U-tube containing pellets of
Nitrous Oxide contains not less than 98.0 per cent v/v of N2O. potassium hydroxide, a U-tube containing phosphorus
NOTE — Carry out the following tests on a full cylinder from pentoxide dispersed on previously granulated, fused pumice,
which no gas has been withdrawn. The cylinder from which a tube containing iodine pentoxide in granules, previously
the gas is taken should be kept at room temperature for not dried at 200° and kept at a temperature of 120°, packed in 1-cm

836
IP 2007 NORADRENALINE BITARTRATE

columns separated by 1-cm columns of glass wool giving an solution of 0.5 g of soluble starch and 0.5 g of potassium
effective length of 5 cm, and a flask containing 2.0 ml of 1 M iodide in 100 ml of water containing 0.05 ml of glacial acetic
potassium iodide and 0.15 ml of starch solution. acid; the colour of the liquid is not changed.
Flush the apparatus with 5.0 litres of carbon dioxide-free air Water. Pass a measured quantity at a rate of 6 litres per hour
and, if necessary, discharge the blue colour in the iodide through an absorption tube containing magnesium
solution by adding a small quantity of freshly prepared perchlorate; the increase in weight of the tube does not exceed
0.002 M sodium thiosulphate. Continue flushing until not 2 mg per litre of gas, the initial and final weighings of the tube
more than 0.045 ml of 0.002 M sodium thiosulphate is required being made when the air in it has been displaced by the nitrous
after passing 5.0 litres of carbon dioxide-free air. Pass 5.0 litres oxide.
of the gas under examination through the apparatus and flush Assay. Carry out the assay of nitrous oxide (2.3.32), using
the last traces of liberated iodine into the reaction flask by 100 ml of the gas under examination. Use a cylinder of the gas
passing through the apparatus 1.0 litre of carbon monoxide- under examination from which at least 1 per cent w/w of the
free air. Titrate the liberated iodine with 0.002 M sodium contents have been removed.
thiosulphate. Carry out a blank titration under the same
Storage. Store under pressure in metal cylinders of the type
conditions, using 5.0 litres of carbon dioxide-free air. The
conforming to the appropriate safety regulations and at a
difference between the volumes of 0.002 M sodium
temperature not exceeding 37°.
thiosulphate used in the two titrations is not greater than
1.0 ml. Labelling. The cylinder is painted blue and carries a label
stating “Nitrous Oxide”. In addition, “Nitrous Oxide” or the
Halogens and hydrogen sulphide. Pass a volume containing symbol “N2O” should be stencilled in paint on the shoulder of
1.0 litre measured at 25° and at 101.3 kPa through a mixture of the cylinder.
100 ml of water and 1 ml of silver nitrate solution; neither
opalescence nor darkening is produced.
Nitric oxide and nitrogen dioxide. Not more than 2 ppm v/v in Noradrenaline Bitartrate
both the liquid and gaseous phases, determined by the
following method. Use two of the cylinders described in the Noradrenaline Acid Tartrate; Levarterenol Bitartrate;
test for Acidity or alkalinity connected in series. Examine Norepinephrine Bitartrate
separately both the liquid and gaseous phases of the gas
OH
under examination. To obtain the liquid phase invert the
cylinder. The liquid vaporises on leaving the valve. NH2 H OH
, HOOC COOH , H2O
For solution A dissolve 1 g of sulphanilic acid in a mixture of
HO
10 ml of glacial acetic acid and 180 ml of water. For solution H OH
B dissolve 0.2 g of N-(1-naphthyl)ethylenediamine OH
dihydrochloride in 10 ml of a 50 per cent v/v solution of
C8H11NO3,C4H6O6,H2O Mol. Wt. 337.3
glacial acetic acid, heating gently, and dilute to 200 ml with
water. Mix 9 volumes of solution A with 1 volume of solution Noradrenaline Bitartrate is (R)-2-amino-1-(3,4-
B (reagent A). dihydroxyphenyl)ethanol hydrogen (2R,3R)-tartrate
monohydrate.
In the first cylinder place 15 ml of a solution containing 2.5 per
cent w/v solution of potassium permanganate and 1.2 per Noradrenaline Bitartrate contains not less than 98.5 per cent
cent v/v of sulphuric acid (96 per cent). Place 20 ml of reagent and not more than 101.0 per cent of C8H11NO3,C4H6O6,
A in the second cylinder and connect the outlet tube of the calculated on the anhydrous basis.
first cylinder to the inlet tube of the second cylinder. Pass Description. A white or almost white, crystalline powder;
2.5 litres of the gas under examination through the reagents at odourless. It gradually darkens on exposure to air and light.
a rate of 15 litres per hour. Prepare a reference solution by
adding 0.25 ml of a 0.00616 per cent w/v solution of sodium Identification
nitrite to 20 ml of reagent A. Allow both the sample and Test A may be omitted if tests B, C, D, E and F are carried out.
reference solutions to stand for 10 minutes. For both liquid Tests C, D, E may be omitted if tests A, B and F are carried out.
and gaseous phases, any red colour in the sample solution is
not more intense than that in the reference solution. A. Dissolve 0.2 g in 2 ml of water containing about 10 mg of
sodium sulphite and add sufficient dilute ammonia solution
Oxidising substances. Pass a volume containing 2.0 litres to give an alkaline reaction. Keep the mixture at about 4° for
measured at 25° and at 101.3 kPa through a freshly prepared 1 hour and filter.

837
NORADRENALINE BITARTRATE INJECTION IP 2007

On the residue (residue R) determine by infrared absorption Apply to the plate 6 µl of each of test solution, reference
spectrophotometry (2,4,6). Compare the spectrum with that solutions (a) and (b) and 12 µl of reference solution (c) as
obtained with noradrenaline acid tartrate RS treated in the bands 20 mm by 2 mm. Allow the applied bands to dry in air,
same manner. spray them with a saturated solution of sodium bicarbonate,
B. When examined in the range 230 nm to 360 nm (2.4.7), a allow to dry in air and spray the bands twice with acetic
0.005 per cent w/v solution in 0.01 M hydrochloric acid shows anhydride, drying between the two sprayings. Heat the plate
an absorption maximum at about 279 nm; absorbance at about at 50° for 90 minutes and develop the chromatograms. After
279 nm, about 0.40. removal of the plate, allow it to dry in air and spray with a
freshly prepared mixture of 8 volumes of methanol, 2 volumes
C. Wash residue R obtained in test A with three quantities, of ethylenediamine and 2 volumes of a 0.5 per cent w/v
each of 2 ml, of water, followed by 5 ml of ethanol (95 per solution of potassium ferricyanide. Dry the plate at 60° for
cent) and 5 ml of ether and dry the precipitate under pressure 10 minutes and examine in ultraviolet light at 254 and 365 nm.
of 1.5 to 2.5 kPa for 3 hours. The specific optical rotation In the chromatogram obtained with the test solution any band
(2.4.22), determined in a 2.0 per cent w/v solution of the dried with a slightly higher Rf value than the principal band is not
precipitate in 0.5 M hydrochloric acid is –44° to –48°. more intense than the corresponding band in the chromatogram
D. To 1 ml of a 1 per cent w/v solution, add 0.05 ml of ferric obtained with reference solution (b). The chromatogram
chloride solution; an intense green colour is produced. Add, obtained with reference solution (c) shows a clearly separated
drop by drop, sodium bicarbonate solution; the colour band corresponding to the most intense band in the
changes to blue and then red. chromatogram obtained with reference solution (a) at a higher
Rf value than the most intense band.
E. To 1 ml of a 0.1 per cent w/v solution add 10 ml of phthalate
buffer pH 3.6, add 1 ml of 0.05 M iodine, set aside for 5 minutes Noradrenalone. Absorbance of a 0.2 per cent w/v solution in
and add 2 ml of 0.1 M sodium thiosulphate; not more than a 0.01 M hydrochloric acid at 310 nm, not more than 0.40 (2.4.7).
faint red colour is produced. Repeat the test using buffer Sulphated ash (2.3.18). Not more than 0.1 per cent.
solution pH 6.6 instead of phthalate buffer pH 3.6; a strong Water (2.3.43). 4.5 to 5.8 per cent, determined on 0.5 g.
reddish violet colour is produced (distinction from adrenaline Assay. Weigh accurately about 0.6 g, dissolve in 50 ml of
and isoprenaline). anhydrous glacial acetic acid, warming if necessary.Titrate
F. The filtrate obtained in test A gives the reactions of tartrates with 0.1 M perchloric acid, using crystal violet solution as
(2.3.1). indicator, until a bluish green colour is obtained. Carry out a
blank titration.
Tests 1 ml of 0.1 M perchloric acid is equivalent to 0.03193 g of
Appearance of solution. A freshly prepared 2.0 per cent w/v C8H11NO3,C4H6O6.
solution is clear (2.4.1), and not more intensely coloured than Storage. Store protected from moisture.
reference solution BYS5 (2.4.1).
pH (2.4.24). 3.5 to 5.0, determined in a 1.0 per cent w/v solution.
Noradrenaline Bitartrate Injection
Melting range (2.4.21). 100° to 106°, with decomposition.
Noradrenaline Acid Tartrate Injection; Noradrenaline
Adrenaline. Determine by thin-layer chromatography (2.4.17),
Injection; Levarterenol Bitartrate Injection;
coating the plate with silica gel G.
Norepinephrine Bitartrate Injection
Mobile phase. A mixture of 50 volumes of acetone, 50 volumes
Noradrenaline Bitartrate Injection is a sterile solution of
of dichloromethane and 0.5 volume of anhydrous formic acid.
Noradrenaline Bitartrate. It is prepared by diluting Sterile
Prepare the following solutions immediately before use. Noradrenaline Concentrate to 250 times its volume with Sodium
Test solution. Dissolve 0.25g of the substance under Chloride and Dextrose Injection or with Dextrose Injection
examination in 10 ml of water. (5 per cent w/v) immediately before use.
Reference solution (a). A 0.125 per cent w/v solution of Noradrenaline Bitartrate Injection contains in 1 ml 8 µg of
adrenaline tartrate RS in water. Noradrenaline Bitartrate equivalent to approximately 4 µg of
noradrenaline.
Reference solution (b). A 0.025 per cent w/v solution of
adrenaline tartrate RS in water. Tests
Reference solution (c). A mixture of equal volumes of the test Other Tests. Complies with the tests stated under Parenteral
solution and reference solution (b). Preparations (Injections).

838
IP 2007 NORETHISTERONE

Sterile Noradrenaline Concentrate Description. A white or yellowish-white, crystalline powder;


Sterile Noradrenaline Concentrate is a sterile, isotonic solution odourless.
containing 0.2 per cent w/v of Noradrenaline Bitartrate in Identification
Water for Injections.
Sterile Noradrenaline Concentrate contains not less than A. Determine by infrared absorption spectrophotometry (2.4.6).
0.18 per cent and not more than 0.23 per cent w/v of Compare the spectrum with that obtained with norethisterone
noradrenaline bitartrate, C8H11NO3,C4H6O6,H2O. RS or with the reference spectrum of norethisterone.
B. Determine by thin-layer chromatography (2.4.17), coating
Identification the plate with silica gel G.
Mix 0.5 ml with 10 ml of phthalate buffer pH 3.6, add 1 ml of Solvent mixture. A mixture of 90 volumes of acetone and
0.05 M iodine, allow to stand for 5 minutes and add 2 ml of 10 volumes of formamide.
0.1 M sodium thiosulphate; not more than a very faint red
colour is produced. Repeat the test using phosphate buffer Mobile phase. A mixture of 40 volumes of hexane and
pH 6.6 instead of phthalate buffer pH 3.6; a strong reddish 10 volumes of dioxan.
violet colour is produced. Test solution. Dissolve 10 mg of the substance under
examination in 10 ml of chloroform.
Tests
Reference solution. Dissolve 10 mg of norethisterone RS in
pH (2.4.24). 3.0 to 4.6.
10 ml of chloroform.
Other Tests. Complies with the tests stated under Parenteral
Preparations (Concentrated Solutions for Injection). Place the dry plate in a tank containing a shallow layer of the
solvent mixture, allow the solvent mixture to ascend to the
Assay. Dilute 5.0 ml to 200.0 ml with water and measure the top, remove the plate from the tank and allow the solvent to
absorbance of the resulting solution at the maximum at about evaporate. Use within 2 hours, with the flow of the mobile
279 nm (2.4.7). Calculate the content of C8H11NO3,C4H6O6,H2O phase in the direction in which the aforementioned treatment
taking 80 as the specific absorbance at 279 nm. was done.
Storage. Store protected from light, in single dose containers.
Apply to the plate 2 µl of each solution. Allow the mobile
Labelling. The label states (1) “Sterile Noradrenaline phase to rise 12 cm. Dry the plate in a current of warm air, allow
Concentrate”; (2) that 1 volume of the solution diluted to the solvent to evaporate, heat at 120° for 15 minutes and spray
250 volumes with Sodium Chloride and Dextrose Injection or the hot plate with ethanolic sulphuric acid (20 per cent v/v).
with Dextrose Injection (5 per cent w/v) produces Heat at 120° for a further 10 minutes, allow to cool and examine
Noradrenaline Bitartrate Injection, which must be used in daylight and in ultraviolet light at 365 nm. The principal
immediately after preparation; (3) that if the solution is brown spot in the chromatogram obtained with the test solution
it should not be used. corresponds to and exhibits fluorescence similar to that in the
chromatogram obtained with the reference solution.
C. Dissolve about 2 mg in 2 ml of ethanol (95 per cent) and
Norethisterone add 1 ml of a 1 per cent w/v solution of butylated
Norethindrone hydroxytoluene in ethanol (95 per cent) and 2 ml of 1 M
sodium hydroxide. Heat in a water-bath for 30 minutes and
cool; a yellowish pink colour is produced.
H3C OH
C CH
Tests
H H
Appearance of solution. Dissolve 0.2 g in sufficient dioxan to
H H produce 10 ml (solution A). The solution is clear (2.4.1), and
O not more intensely coloured than reference solution YS6 (2.4.1).
C20H26O2 Mol. Wt. 298.4 Specific optical rotation (2.4.22). –33.0° to –37.0°, determined
Norethisterone is 17β-hydroxy-19-nor-17α-pregn-4-en-20-yn- in a solution prepared by diluting 5.0 ml of solution A to
3-one. 10.0 ml with dioxan.
Norethisterone contains not less than 98.0 per cent and not Light absorption. Dissolve 10 mg in sufficient ethanol
more than 102.0 per cent of C20H26O2, calculated on the dried (95 per cent) to produce 100 ml, dilute 10 ml of the solution to
basis. 100 ml with methanol (98 per cent). Absorbance of the

839
NORETHISTERONE TABLETS IP 2007

resulting solution at the maximum at about 240 nm, 0.55 to 0.59 each of 5 ml, of light petroleum (60° to 80°) and discard the
(2.4.7). washings. Extract the residue with 15 ml of chloroform,
Related substances. Determine by thin-layer chromatography evaporate the extract to dryness and recrystallise from aqueous
(2.4.17), coating the plate with silica gel GF254. methanol. The residue complies with the following test.

Mobile phase. A mixture of 90 volumes of chloroform and Determine by thin-layer chromatography (2.4.17), coating the
10 volumes of acetone. plate with silica gel G.

Test solution. Dissolve 0.5 g of the substance under Solvent mixture. A mixture of 90 volumes of acetone and
examination in 100 ml of the mobile phase. 10 volumes of 1,2-propanediol.
Reference solution (a). A 0.0025 per cent w/v solution of the Mobile phase. A mixture of 40 volumes of cyclohexane and 10
substance under examination in the mobile phase. volumes of toluene.
Reference solution (b). A solution containing 0.025 per cent Test solution. Dissolve 25 mg of the substance under
w/v each of the substance under examination and ethisterone examination in 10 ml of the solvent mixture.
RS in the mobile phase.
Reference solution (a). Dissolve 25 mg of norethisterone RS
Apply to the plate 10 µl of each solution. After development, in 10 ml of the solvent mixture.
dry the plate in air, spray with ethanolic sulphuric acid
Reference solution (b). Mix equal volumes of the test solution
(20 per cent v/v), heat at 105° for 5 minutes and examine in
and reference solution (a).
ultraviolet light at 365 nm. Any secondary spot in the
chromatogram obtained with the test solution is not more Place the dry plate in a tank containing a shallow layer of the
intense than the spot in the chromatogram obtained with solvent mixture, allow the solvent mixture to ascend to the
reference solution (a). The chromatogram obtained with top, remove the plate from the tank and allow the solvent to
reference solution (b) shows two clearly separated spots of evaporate. Use within 2 hours, with the flow of the mobile
equal intensities. phase in the direction in which the aforementioned treatment
Sulphated ash (2.3.18). Not more than 0.1 per cent. was done.

Loss on drying (2.4.19). Not more than 0.5 per cent, determined Apply to the plate 2 µl of each solution. Allow the mobile
on 1.0 g by drying in an oven at 105° for 3 hours. phase to rise 12 cm. Dry the plate in a current of warm air, allow
the solvent to evaporate, heat at 120° for 15 minutes and spray
Assay. Weigh accurately about 0.4 g, dissolve in 40 ml of the hot plate with ethanolic sulphuric acid (20 per cent v/v).
tetrahydrofuran, add 10 ml of a 10 per cent w/v solution of Heat at 120° for a further 10 minutes, allow to cool and examine
silver nitrate and titrate with 0.1 M sodium hydroxide using in daylight and in ultraviolet light at 365 nm. The principal
2 ml of bromocresol green solution as indicator. Repeat the spot in the chromatogram obtained with the test solution
operation without the substance under examination. The corresponds to that in the chromatogram obtained with
difference between the titrations represents the amount of reference solution (a). The principal spot in the chromatogram
sodium hydroxide required. obtained with reference solution (b) appears as a single,
1 ml of 0.1 M sodium hydroxide is equivalent to 0.02984 g of compact spot.
C20H26O2.
Tests
Storage. Store protected from light and moisture.
Uniformity of content. Comply with the test stated under
Tablets.
Norethisterone Tablets Powder one tablet and warm with about 75 ml of ethanol
(95 per cent) with stirring. Cool, transfer to a 100-ml volumetric
Norethindrone Tablets
flask and dilute to volume with ethanol (95 per cent).
Norethisterone Tablets contain not less than 90.0 per cent Centrifuge a few ml of the mixture until a clear supernatant
and not more than 110.0 per cent of the stated amount of liquid is obtained. Dilute 10.0 ml of the supernatant liquid to
norethisterone, C20H26O2. 50.0 ml with ethanol (95 per cent) and mix. Measure the
absorbance of the resulting solution at the maximum at about
Identification 240 nm (2.4.7). Calculate the content of C20H26O2 in the tablet
taking 570 as the specific absorbance at 240 nm.
Place a quantity of the powdered tablets containing 25 mg of
Norethisterone on a small filter, wash with three quantities, Other Tests. Comply with the tests stated under Tablets.

840
IP 2007 NORFLOXACIN

Assay. Weigh and powder 20 tablets. Weigh accurately a Tests


quantity of the powder containing about 0.2 g of
Norethisterone and transfer to a glass column closed at the Related substances. Determine by thin-layer chromatography
bottom with a small piece of absorbent cotton. The glass (2.4.17), coating the plate with silica gel G, previously washed
column consists of a piece of glass tubing (150 mm x 10 mm) with methanol and dried.
tapered at the bottom and sealed at the top to another piece of Mobile phase. A mixture of 40 volumes of dichloromethane,
glass tubing (150 mm x 25 mm). Place a small piece of absorbent 40 volumes of methanol, 20 volumes of toluene, 14 volumes
cotton on top of the powder, pass 200 ml of light petroleum of diethylamine and 8 volumes of water.
(60° to 80°) through the column and discard the effluent.
Extract the residue with 200 ml of chloroform, evaporate the Test solution. Dissolve 0.8 g of the substance under
chloroform from the extract and dry the residue at 105° for examination in 100 ml of a mixture of equal volumes of methanol
2 hours. Allow to cool, dissolve in 40 ml of tetrahydrofuran, and dichloromethane.
add 10 ml of a 10 per cent w/v solution of silver nitrate. Titrate Reference solution. Dissolve 4.0 mg of norfloxacin RS in 1 ml
with 0.1 M sodium hydroxide, determining the end-point of glacial acetic acid, add 4 ml of methanol and mix; dilute
potentiometrically (2.4.25). Repeat the operation without the 1 ml of the solution with 9 ml of a mixture of equal volumes of
substance under examination. The difference between the methanol and dichloromethane (reference solution A). Dilute
titrations represents the amount of sodium hydroxide required. a portion of reference solution A with an equal volume of the
1 ml of 0.1 M sodium hydroxide is equivalent to 0.02984 g of methanol-dichloromethane mixture (reference solution B).
C20H26O2. Apply separately to the plate spots of the three solutions in
Storage. Store protected from light and moisture. quantities indicated below. For spot 1 use 5 µl of the test
solution; for spots 2, 3 and 4 use 1 µl, 1.5 µl and 2 µl respectively
of reference solution A; for spot 5 use 5 µl of reference solution
B. Place the plate in a paper-lined chamber previously
equilibrated with the mobile phase and allow the solvent front
Norfloxacin to move about nine-tenths of the length of the plate. After
development, dry the plate in air and examine in ultraviolet
light at 254 nm and 365 nm. Compare the intensities of any
CH3 secondary spots in the chromatogram obtained with the test
HN
solution with those of the principal spots (2), (3), (4) and (5).
N N The sum of the intensities of secondary spots obtained with
the test solution is not more than 0.5 per cent of impurities.
F COOH (The spots (2) (3) (4) and (5) represent 0.2 per cent, 0.3 per
O cent, 0.4 per cent and 0.5 per cent respectively of impurities).
Heavy metals (2.3.13). 1.33 g complies with the limit test for
C16H18FN3O3 Mol. Wt. 319.3 heavy metals, Method B (15 ppm).

Norfloxacin is 1-ethyl-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4- Sulphated ash (2.3.18). Not more than 0.1 per cent, determined
dihydroquinoline-3-carboxylic acid. on 1.0 g in a platinum crucible.
Loss on drying (2.4.19). Not more than 1.0 per cent, determined
Norfloxacin contains not less than 99.0 per cent and not more
on 0.5 g by drying in an oven at 100° at a pressure not exceeding
than 101.0 per cent of C16H18FN3O3, calculated on the dried
0.7 kPa.
basis.
Assay. Weigh accurately about 0.3 g and dissolve in 100 ml of
Description. A white to pale yellow, crystalline powder.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
acid, determining the end-point potentiometrically (2.4.25) and
Identification
using a suitable anhydrous electrode system. (The electrode
A. Determine by infrared absorption spectrophotometry (2.4.6). system may be rendered anhydrous by filling the electrode
Compare the spectrum with that obtained with norfloxacin with 0.1 M lithium perchlorate in acetic anhydride after
RS or with the reference spectrum of norfloxacin. removing any aqueous solution contained in it). Carry out a
blank titration.
B. When examined in the range 230 nm to 360 nm (2.4.7), a
0.0005 per cent w/v solution in 0.1 M sodium hydroxide shows 1 ml of 0.1 M perchloric acid is equivalent to 0.03193 g of
an absorption maximum at about 273 nm. C16H18FN3O3.

841
NORFLOXACIN EYE DROPS IP 2007

Storage. Store protected from light and moisture. Norfloxacin Tablets


Norfloxacin Tablets contain not less than 90.0 per cent and
not more than 110.0 per cent of the stated amount of
Norfloxacin Eye Drops norfloxacin, C16H18FN3O3. The tablets may be coated.
Norfloxacin Eye Drops are a sterile solution of Norfloxacin in
Purified water.
Identification

Norfloxacin Eye Drops contain not less than 90.0 per cent and A. Determine by thin-layer chromatography (2.4.17), coating
not more than 110.0 per cent of the stated amount of the plate with silica gel GF254.
norfloxacin, C16H18FN3O3. Mobile phase. A mixture of 40 volumes of chloroform,
40 volumes of methanol, 20 volumes of toluene, 14 volumes
Identification of diethylamine and 8 volumes of water.
In the Assay, the principal peak in the chromatogram obtained Test solution. Shake a quantity of the finely powdered tablets
with the test solution corresponds to the peak in the containing 75 mg of Norfloxacin with 50 ml of a mixture of
chromatogram obtained with the reference solution. equal volumes of acidified methanol (containing 0.9 per cent
v/v of hydrochloric acid) and dichloromethane, centrifuge
Tests and use the clear supernatant solution.
pH (2.4.24). 4.6 to 5.5. Reference solution. A 0.15 per cent w/v solution of norfloxacin
Other tests. Comply with the tests stated under Eye Drops. RS in the same solvent mixture.

Assay. Determine by liquid chromatography (2.4.14). Apply to the plate 50 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
Test solution. Dilute a suitable volume of the eye drops with a The principal spot in the chromatogram obtained with the test
0.1 per cent v/v solution of orthophosphoric acid to produce solution corresponds to that in the chromatogram obtained
a solution containing 0.005 per cent w/v of Norfloxacin. with the reference solution.
Reference solution. A 0.005 per cent w/v solution of B. In the Assay, the principal peak in the chromatogram
norfloxacin RS in 0.1 per cent v/v solution of orthophosphoric obtained with the test solution corresponds to the peak due
acid. to norfloxacin in the chromatogram obtained with the reference
Chromatographic system solution.
– a stainless steel column 30 cm x 3.9 mm, packed with
octadecylsilyl silica gel (10 µm) (such as Bondapack Tests
C18), Dissolution (2.5.2).
– column temerature 50°,
Apparatus. No 1
– mobile phase: a mixture of 300 volumes of methanol
Medium. 750 ml of acetate buffer pH 4.0.
and 700 volumes of 0.1 per cent v/v orthophosphoric
acid, Speed and time. 50 rpm and 30 minutes.
– flow rate. 2 ml per minute, Withdraw a suitable volume of the medium and filter. Measure
– spectrophotometer set at 280 nm, the absorbance of the filtrate, suitably diluted with acetate
– a 20 µl loop injector. buffer pH 4.0, if necessary, at the maximum at about 278 nm
Precondition the column using 0.01 M anhydrous sodium (2.4.7). Concomitantly measure the absorbance of a solution
dihydrogen orthophosphate, adjusted to pH 4.0 with of known concentration of norfloxacin RS in the same medium.
orthophosphoric acid, at a flow rate of 0.5 ml per minute for 8 Calculate the total content of C16H18FN3O3 in the medium.
hours. Equilibrate the column with the mobile phase for about D. Not less than 70 per cent of the stated amount of
30 minutes before starting the chromatography. C16H18FN3O3.
Inject the reference solution. The test is not valid unless the
Other Tests. Comply with the tests stated under Tablets.
tailing factor is not more than 2.0. The relative standard
deviation for replicate injections is not more than 2.0 per cent. Assay. Determine by liquid chromatography (2.4.14).
Inject alternately the test solution and the reference solution. Test solution. Weigh and powder 20 tablets. Add 80 ml of the
mobile phase to an accurately weighed quantity of the
Calculate the content of C16H18FN3O3 in the eye drops. powdered tablets containing about 100 mg of Norfloxacin, mix
Storage. Store protected from light. with the aid of ultrasound for 10 minutes, dilute with a 0.1 per

842
IP 2007 NORGESTREL AND ETHINYLOESTRADIOL TABLETS

cent v/v solution of phosphoric acid to 200.0 ml and mix. C. Melting range (2.4.21). 205° to 212°, but the range between
Dilute 10.0 ml of this solution to 25.0 ml with the mobile phase, beginning and end of melting does not exceed 4°.
mix and use the resulting solution after filtration through a
filter with porosity of not more than 0.1 µm. Tests
Reference solution. A 0.02 per cent w/v solution of norfloxacin Specific optical rotation (2.4.22). –0.1° to +0.1°, determined in
RS in the mobile phase. a 5.0 per cent w/v solution in chloroform.
Chromatographic system Related substances. Determine by thin-layer chromatography
– a stainless steel column 30 cm x 3.9 mm, packed with (2.4.17), coating the plate with silica gel G.
octadecylsilane bonded to porous silica (5 µm), Mobile phase. A mixture of 80 volumes of dichloromethane
– mobile phase: 85 volumes of a 0.1 per cent v/v solution and 20 volumes of ethyl acetate.
of phosphoric acid and 15 volumes of acetonitrile,
– temperature. column 40° ± 1°, after preconditioning with Test solution. Dissolve 0.2 g of the substance under
degassed 0.01 M sodium dihydrogen phosphate examination in 10 ml of chloroform.
adjusted to pH 4.0 with phosphoric acid flowing at a Reference solution (a). A 0.01 per cent w/v solution of the
rate of 0.5 ml per minute for 8 hours, substance under examination in chloroform.
– flow rate. 2 ml per minute,
Reference solution (b). A 0.004 per cent w/v solution of the
– spectrophotometer set at 275 nm,
substance under examination in chloroform.
– a 20 µl loop injector.
Apply to the plate 10 µl of each solution. After development,
Inject the test solution and the reference solution. The assay
dry the plate in air and spray with phosphomolybdic acid
is not valid unless the capacity factor is not less than 2, the
solution. Any secondary spot in the chromatogram obtained
column efficiency is not less than 1500 theoretical plates, the
with the test solution is not more intense than the spot in the
tailing factor for the norfloxacin peak is not more than 2.0 and
chromatogram obtained with reference solution (a) and not
the relative standard deviation for replicate injections is not
more than two such spots are more intense than the spot in
more than 2.0 per cent.
the chromatogram obtained with reference solution (b).
Calculate the content of C16H18FN3O3 in the tablets. Sulphated ash. (2.3.18) Not more than 0.3 per cent.
Storage. Store protected from light and moisture. Loss on drying (2.4.19). Not more than 0.5 per cent, determined
on 1.0 g by drying in an oven at 105° for 5 hours.
Assay. Weigh accurately about 0.1 g, dissolve in sufficient
Norgestrel ethanol (95 per cent) to produce 100.0 ml, dilute stepwise
with ethanol (95 per cent) to obtain a solution containing
H3 C OH 0.001 per cent w/v of Levonorgestrel and measure the
C CH absorbance of the resulting solution at the maximum at about
H H 241 nm, (2.4.7). Calculate the content of C21H28O2 from the
absorbance obtained with a 0.001 per cent w/v solution of
H H norgestrel RS in ethanol (95 per cent).
O Storage. Store protected from moisture.
C21H28O2 Mol. Wt. 312.5
Norgestrel is rac-13-ethyl-17-hydroxy-18,19-dinor-17α-
pregn-4-en-20-yn-3-one. Norgestrel and Ethinyloestradiol
Description. A white or almost white, crystalline powder; Tablets
practically odourless. Norgestrel and Ethinyloestradiol Tablets contain not less than
90.0 per cent and not more than 110.0 per cent of the stated
Identification
amounts of norgestrel, C21H28O 2 and ethinyloestradiol,
A. Determine by infrared absorption spectrophotometry (2.4.6). C20H24O2. The tablets may be film-coated.
Compare the spectrum with that obtained with norgestrel RS.
Identification
B. When examined in the range 220 nm to 360 nm (2.4.7), a
0.001 per cent w/v solution in methanol shows an absorption Determine by thin-layer chromatography (2.4.17), coating the
maximum only at about 240 nm. plate with silica gel G.

843
NORTRIPTYLINE HYDROCHLORIDE IP 2007

Mobile phase. A mixture of 96 volumes of dichloromethane – mobile phase: a mixture of 35 volumes of acetonitrile,
and 4 volumes of ethanol (95 per cent). 15 volumes of methanol and 45 volumes of water,
Test solution. Powder 20 tablets finely, triturate with 20 ml of – flow rate. 1 to1.5 ml per minute,
dichloromethane, allow the solids to sediment and use the – spectrophotometer set at 215 nm,
clear supernatant liquid. – a 20 µl loop injector.

Reference solution. A solution containing 0.06 per cent w/v Inject the reference solution. The resolution between the two
solution of norgestrel RS and 0.006 per cent w/v solution of major peaks is not less 2.5 and the relative standard deviation
ethinyloestradiol RS. for replicate injections is not more than 2.0 per cent.

Apply to the plate 40 µl of each solution. After development, Inject the test solution and the reference solution. The relative
dry the plate in air, spray with ethanolic sulphuric acid retention times are about 0.7 for ethinyloestradiol and about
(80 per cent v/v), heat at 110° for 10 minutes and examine in 1.0 for norgestrel.
ultraviolet light at 365 nm. The principal spots in the Calculate the contents of norgestrel, C 21 H 28 O 2 , and
chromatogram obtained with the test solution correspond to ethinyloestradiol, C20H24O2 in the tablets.
the spots for norgestrel (red fluorescence) and
Storage. Store protected from light.
ethinyloestradiol (orange-yellow fluorescence) in the
chromatogram obtained with the reference solution.

Tests
Nortriptyline Hydrochloride
Uniformity of content. Comply with the test stated under
Tablets.
Carry out the procedure described under Assay but using the
following test solution. , HCl
Test solution. Add 2.0 ml of methanol (70 per cent) and 2.0 ml CH3
of a 0.00002 per cent w/v solution of diphenyl in methanol N
H
(70 per cent) (internal standard solution) to one tablet, shake
for 20 minutes, centrifuge, filter the supernatant liquid through C19H21N,HCl Mol. Wt. 299.8
a membrane filter with a pore size of not more than 0.2 mm and
Nortriptyline Hydrochloride is 3-(10,11-dihydro-5H-dibenzo
use the filtrate.
[a,d]cyclohept-5-ylidene)propyl(methyl)amine hydrochloride.
Calculate the contents of norgestrel C 21 H 28 O 2 , and
Nortriptyline Hydrochloride contains not less than 98.0 per
ethinyloestradiol, C20H24O2, in the tablet.
cent and not more than 101.5 per cent of C19H21N,HCl, calculated
Other Tests. Comply with the tests stated under Tablets. on the dried basis.
Assay. Determine by liquid chromatography (2.4.14) using the Description. A white to off-white powder; odour slight and
chromatographic system described under Uniformity of characteristic.
content.
Test solution. Weigh and powder 20 tablets. To a quantity of Identification
the powder equivalent to one tablet add 2.0 ml of methanol Test A may be omitted if tests B, C and D are carried out. Tests
(70 per cent) and 2.0 ml of a 0.00002 per cent w/v solution of B and C may be omitted if tests A and, D are carried out.
diphenyl in methanol (70 per cent) (internal standard
solution), shake for 20 minutes, centrifuge, filter the A. Dissolve 0.1 g in 10 ml of water, make alkaline with 1 M
supernatant liquid through a membrane filter with a pore size sodium hydroxide, extract with 5 ml of chloroform and
of not more than 0.2 mm and use the filtrate. evaporate to dryness using a current of nitrogen.
Reference solution. A solution in methanol (70 per cent ) Determine by infrared absorption spectrophotometry (2.4.6).
containing 0.15 mg per ml of norgestrel RS and 0.015 mg per Compare the spectrum with that obtained with nortriptyline
ml of ethinyloestradiol RS. Take 2.0 ml of this solution and hyrdochloride RS treated in the same manner or with the
add 2.0 ml of a 0.00002 per cent w/v solution of diphenyl in reference spectrum of nortriptyline.
methanol (70 per cent) and use the resulting solution. B. When examined in the range 230 nm to 360 nm (2.4.7), a
Chromatographic system 0.001 per cent w/v solution in methanol shows an absorption
– a stainless steel column 15 cm x 4.6 mm, packed with maximum only at about 239 nm; absorbance at about 239 nm,
octadecylsilane bonded to porous silica (5 to 7 µm), about 0.48.

844
IP 2007 NORTRIPTYLINE TABLETS

C. To about 50 mg dissolved in 3 ml of warm water, add 1 drop Identification


of a 2.5 per cent w/v solution of quinhydrone in methanol; a
red colour is produced after a few minutes (distinction from A. Shake a quantity of the powdered tablets containing about
amitriptyline). 5 mg of nortriptyline with 20 ml of methanol and filter. To 1 ml
of the filtrate add 1 ml of a 2.5 per cent w/v solution of sodium
D. Gives the reactions of chlorides (2.3.1). bicarbonate, 1 ml of a 2 per cent w/v solution of sodium
periodate and 1 ml of a 0.3 per cent w/v solution of potassium
Tests permanganate. Allow to stand for 15 minutes, acidify with
Related substances. Determine by thin-layer chromatography 1 M sulphuric acid and extract with 10 ml of 2,2,4-
(2.4.17), coating the plate with silica gel G. trimethylpentane.

Mobile phase. A mixture of 85 volumes of cyclohexane, When examined in the range 230 nm to 360 nm (2.4.7), the
15 volumes of ethyl acetate and 3 volumes of diethylamine. resulting trimethylpentane solution shows an absorption
maximum only at about 265 nm.
Test solution. Dissolve 0.2 g of the substance under
B. Triturate a quantity of the powdered tablets containing
examination in 10 ml of ethanol (95 per cent) prepared in
0.1 g of nortriptyline with 10 ml of chloroform, filter and
subdued light.
evaporate the filtrate to a low volume. Add ether until a
Reference solution. A 0.001 per cent w/v solution of turbidity is produced and allow to stand. Dissolve 50 mg of
dibenzosuberone RS in ethanol (95 per cent) prepared in the precipitate in 3 ml of warm water, cool and add 1 drop of a
subdued light. 2.5 per cent w/v solution of quinhydrone in methanol; a red
Apply to the plate 5 µl of each solution. Allow the mobile colour is produced after a few minutes (distinction from
phase to rise 14 cm in an unsaturated tank protected from amitriptyline).
light. Dry the plate in air, spray with a freshly prepared solution
of sulphuric acid containing 4 per cent v/v of formaldehyde
Tests
solution and examine immediately in ultraviolet light at Related substances. Determine by thin-layer chromatography
365 nm. Any secondary spot in the chromatogram obtained (2.4.17), coating the plate with silica gel G.
with the test solution is not more intense than the spot in the
chromatogram obtained with the reference solution. Mobile phase. A mixture of 85 volumes of cyclohexane,
15 volumes of ethyl acetate and 3 volumes of diethylamine.
Heavy metals (2.3.13). 2.0 g complies with the limit test for
Test solution. Extract a quantity of the powdered tablets
heavy metals, Method B (10 ppm).
containing 20 mg of nortriptyline with 5 ml of a mixture of
Sulphated ash (2.3.18). Not more than 0.1 per cent. 9 volumes of ethanol (95 per cent) and 1 volume of 2 M
Loss on drying (2.4.19). Not more than 0.5 per cent, determined hydrochloric acid, centrifuge and use the supernatant liquid.
on 1.0 g by drying in an oven at 105° for 3 hours. Reference solution. A 0.001 per cent w/v solution of
Assay. Weigh accurately about 0.25 g and dissolve in 25 ml of dibenzosuberone RS in ethanol (95 per cent) prepared in
anhydrous glacial acetic acid, warm slightly, if necessary, to subdued light.
effect solution. Cool, add 5 ml of mercuric acetate solution. Apply to the plate 5 µl of each solution. Allow the mobile
Titrate with 0.1 M perchloric acid, determining the end-point phase to rise 14 cm in an unsaturated tank protected from
potentiometrically (2.4.25). Carry out a blank titration. light. Dry the plate in air, spray with a freshly prepared solution
of sulphuric acid containing 4 per cent v/v of formaldehyde
1 ml of 0.1 M perchloric acid is equivalent to 0.02998 g of
solution and examine immediately in ultraviolet light at
C19H21N,HCl.
365 nm. Any secondary spot in the chromatogram obtained
Storage. Store protected from light and moisture. with the test solution is not more intense than the spot in the
chromatogram obtained with the reference solution.
Uniformity of content (For tablets containing 10 mg or less).
Comply with the test stated under Tablets.
Nortriptyline Tablets
Determine by liquid chromatography (2.4.14)
Nortriptyline HydrochlorideTablets
Test solution. Powder one tablet, add 2.5 ml of water, shake
Nortriptyline Tablets contain less than 90.0 per cent and not vigorously to completely disperse the tablet, add 5 ml of
more than 110.0 per cent of the stated amount of nortriptyline, methanol and shake for 30 minutes. Add sufficient water to
C19H21N. The tablets are coated. produce 10 ml, centrifuge and use the clear supernatant liquid.

845
NOSCAPINE IP 2007

Reference solution. A 0.01 per cent w/v solution of Noscapine contains not less than 98.5 per cent and not more
nortriptyline hydrochloride RS in methanol (50 per cent). than 100.5 per cent of C22H23NO7, calculated on the dried basis.
Follow the procedure given in the Assay. Calculate the content Description. Colourless crystals or a white crystalline powder.
of C20H23N in the tablet.
Identification
Other Tests. Comply with the tests stated under Tablets.
Assay. Determine by liquid chromatography (2.4.14). Test A may be omitted if tests B, C and D are carried out. Tests
B, C and D may be omitted if test A is carried out.
Test solution. Shake vigorously 20 tablets with 50 ml of water
until the tablets disintegrate completely, add 100.0 ml of A. Determine by infrared absorption spectrophotometry (2.4.6).
methanol and shake for 30 minutes. Add sufficient water to Compare the spectrum with that obtained with noscapine RS
produce 200.0 ml, filter and dilute a volume of the filtrate or with the reference spectrum of noscapine.
containing about 25 mg of nortriptyline to 100.0 ml with B. When examined in the range 230 nm to 360 nm (2.4.7), a
methanol (50 per cent). 0.005 per cent w/v solution in methanol shows absorption
Reference solution. A 0.025 per cent w/v solution of maxima at about 291 nm and 310 nm; ratio of absorbance at the
nortriptyline hydrochloride RS in methanol (50 per cent). maximum at about 310 nm to that at the maximum at about
291 nm, 1.2 to 1.3.
Chromatographic system
– a stainless steel column 20 cm x 4.6 mm, packed with C. To 0.1 g in a porcelain dish add a few drops of sulphuric
octadecylsilane bonded to porous silica (10 µm), acid and stir; a greenish-yellow solution is formed which on
– mobile phase: a 0.56 per cent w/v solution of sodium warming becomes red and finally violet.
hexanesulphonate in a mixture of equal volumes of D. Dissolve 50 mg in 5 ml of 5 M hydrochloric acid, add 10 ml
water and acetonitrile adjusted to pH 4.5 with glacial of a mixture of equal volumes of ethanol (95 per cent) and a
acetic acid, saturated solution of sodium acetate, mix and allow to stand
– flow rate. 2 ml per minute, for about 3 minutes; shining crystals separate.
– spectrophotometer set at 239 nm,
– a 20 µl loop injector. Tests
Inject the test solution and the reference solution. Appearance of solution. A 2.0 per cent w/v solution in acetone
Calculate the content of C19H21N in the tablets. examined immediately after preparation is clear (2.4.1), and not
more intensely coloured than reference solution YS6 (2.4.1).
Storage. Store protected from light and moisture.
Specific optical rotation (2.4.22). +42.0° to +48.0°, determined
at 20° in a solution prepared by dissolving 0.5 g in sufficient
0.1 M hydrochloric acid to produce 25.0 ml.
Noscapine Related substances. Determine by thin-layer chromatography
Narcotine (2.4.17), coating the plate with silica gel G.
Mobile phase. A mixture of 60 volumes of acetone, 60 volumes
O of toluene, 9 volumes of ethanol (95 per cent) and 3 volumes
of strong ammonia solution.
O N
CH3 Test solution. Dissolve 0.25 g of the substance under
H
OCH3 examination in 10 ml of acetone.
H Reference solution. A 0.0125 per cent w/v solution of the
substance under examination in acetone.
O
Apply to the plate 10 µl of each solution. After development,
H3CO dry the plate in air, spray with dilute potassium
OCH3 O iodobismuthate solution. Any secondary spot in the
chromatogram obtained with the test solution is not more
C22H23NO7 Mol. Wt. 413.4 intense than the spot in the chromatogram obtained with the
reference solution.
Noscapine is (3S)-6,7-dimethoxy-3-[(5R)-5,6,7,8-tetrahydro-
4-methoxy-6-methyl-1,3-dioxolo[4,5-g]isoquinolin-5- Morphine. Dissolve 0.1 g in 10 ml of 0.1 M hydrochloric acid.
yl]phthalide, an alkaloid obtained from opium. To 1 ml of the resulting solution add a mixture of 1 ml of

846
IP 2007 NOVOBIOCIN SODIUM

potassium ferricyanide solution, 0.05 ml of ferric chloride Assay. Weigh accurately a quantity containing 60 mg of
test solution and 4 ml of water; no blue or dark green colour Noscapine, add 20 ml of water, 2 g of sodium chloride and
develops within 1 minute. 2 ml of 5 M sodium hydroxide and extract with successive
Sulphated ash (2.3.18). Not more than 0.1 per cent. quantities of 50, 50, 25 and 25 ml of ether. Combine the extracts,
wash with three quantities, each of 5 ml, of water and evaporate
Loss on drying (2.4.19). Not more than 1.0 per cent, determined to dryness. To the residue add 50.0 ml of 0.1 M hydrochloric
on 1.0 g by drying in an oven at 105°. acid, warm on a water-bath to dissolve and to remove any
Assay. Weigh accurately about 0.35 g, dissolve in 40 ml of traces of ether and dilute to 100.0 ml with water. Dilute 3.0 ml
anhydrous glacial acetic acid, warming gently. Titrate with to 50.0 ml with water and measure the absorbance of the
0.1 M perchloric acid, determining the end-point resulting solution at the maximum at about 310 nm (2.4.7).
potentiometrically (2.4.25). Carry out a blank titration. Calculate the content of C22H23NO7 taking 90.7 as the specific
absorbance at 310 nm.
1 ml of 0.1 M perchloric acid is equivalent to 0.04134 g of
C22H23NO7. Determine the weight per ml of the linctus (2.4.29), and calculate
the content of C22H23NO7, weight in volume.
Storage. Store protected from light and moisture.
Storage. Store protected from light and moisture.

Noscapine Linctus Novobiocin Sodium


Narcotine Linctus CH3
CH3
Noscapine Linctus is a solution of Noscapine in a suitable H3CO O O O
O O CH3
flavoured vehicle. It may contain up to 1 per cent w/v solution CH3
of Citric Acid. N CH3
H3 C O OH
Noscapine Linctus contains not less than 90.0 per cent and ONa H
OH
not more than 110.0 per cent of the stated amount of O
noscapine, C22H23NO7. C31H35N2NaO11 Mol. Wt. 634.6

Identification Novobiocin Sodium is the monosodium salt of novobiocin,


N-[7-{3-O-(aminocarbonyl)-6-deoxy-5-C-methyl-4-O-methyl-
To a quantity containing 60 mg of Noscapine add 20 ml of β-L-lyxo-hexopyranosyl}-4-hydroxy-8-methyl-2-oxo-2H-1-
water, 2 g of sodium chloride and 2 ml of 5 M sodium benzopyran-3-yl]-4-hydroxy-3-(3-methyl-2-
hydroxide. Extract with successive quantities of 50, 50, 25 and butenyl)benzamide, an antimicrobial substance produced by
25 ml of ether. Combine the extracts, wash with three quantities, the growth of certain strains of Streptomyces niveus or
each of 5 ml, of water and evaporate to dryness. Dissolve the related organisms or by other means.
residue in 20 ml of chloroform. Wash with three quantities,
Novobiocin Sodium contains the equivalent of not less than
each of 20 ml, of water, dry the chloroform layer with anhydrous
850 µg of novobiocin per mg, calculated on the dried basis.
sodium sulphate, filter and evaporate the solvent. If necessary,
induce crystallisation by scratching with a glass rod. The Description. A white or yellowish white, crystalline powder.
crystals comply with the following tests.
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with noscapine RS A. Determine by thin-layer chromatography (2.4.17), coating
or with the reference spectrum of noscapine. the plate with silica gel G.
B. When examined in the range 230 nm to 360 nm (2.4.7), a Mobile phase. A mixture of 75 volumes of chloroform,
0.005 per cent w/v solution in methanol shows absorption 25 volumes of methanol and 1 volume of strong ammonia
maxima at about 291 nm and 310 nm; ratio of absorbance at the solution.
maximum at about 310 nm to that at the maximum at about Test solution. Dissolve a quantity of the substance under
291 nm, 1.2 to 1.3. examination in methanol so as to obtain a solution containing
0.1 per cent w/v solution of novobiocin.
Tests
Reference solution. A 0.1 per cent w/v solution of novobiocin
Other tests. Complies with the tests stated under Oral Liquids. RS in methanol.

847
NYSTATIN IP 2007

Apply to the plate 1 µl of each solution. After development, Identification


dry the plate in air and examine in ultraviolet light at 254 nm.
The principal spot in the chromatogram obtained with the test A. When examined in the range 220 nm to 360 nm (2.4.7), the
solution corresponds to that in the chromatogram obtained final solution obtained in the test for Light absorption shows
with the reference solution. absorption maxima at about 230 nm, 291 nm, 305 nm and
319 nm. The ratios of the absorbances at the maxima at about
B. When examined in the range 230 nm to 360 nm (2.4.7), a 291 nm and about 319 nm to the absorbance at the maximum at
0.001 per cent w/v solution in a 0.4 per cent w/v solution of about 305 nm are 0.61 to 0.73 and 0.83 to 0.96, respectively.
potassium hydroxide shows an absorption maximum only at Use as the blank a solution prepared in the same manner
about 307 nm. without the substance under examination.
C. The residue obtained by igniting it gives the tests for sodium
B. Shake 30 mg with 5 ml of water for 2 minutes, add 2 ml of
salts (2.3.1).
sodium molybdotungstophosphate solution, and allow to
Tests stand for 1 hour; the green colour produced is darker than
that produced by repeating the test without the substance
pH (2.4.24). 6.6 to 8.5, determined in a 2.5 per cent w/v solution. under examination.
Specific optical rotation (2.4.22). –50.0° to –58.0°, determined C. Shake 30 mg with 5 ml of water for 2 minutes, add 2 ml of a
in a 5.0 per cent w/v solution in methanol containing 1 per solution prepared by dissolving 0.1 g of pyrogallol in 100 ml
cent v/v of hydrochloric acid. of decolorised magenta solution, heat on a water-bath until a
Loss on drying (2.4.19). Not more than 6 per cent, determined dark pink colour is produced, cool and allow to stand for
on 0.2 g by drying in an oven at 60° over phosphorus pentoxide 1 hour; the pink colour is retained.
at a pressure not exceeding 0.7 kPa for 3 hours. D. To 2 mg add 0.1 ml of hydrochloric acid; a brown colour is
Assay. Determine by the microbiological assay of antibiotics, produced.
Method A (2.2.10), and express the results in µg of novobiocin E. To 2 mg add 0.1 ml of sulphuric acid; a brown colour is
per mg. produced which becomes violet on standing.
Novobiocin Sodium intended for use in the manufacture of
Parenteral Preparations complies with the following Tests
additional requirements.
pH (2.4.24). 6.5 to 8.0, determined in a 3.0 per cent w/v
Bacterial endotoxins (2.2.3). Not more than 0.7 Endotoxin units suspension in water.
per mg.
Light absorption (2.4.7). Dissolve 0.1 g in a mixture of 5.0 ml of
Sterility. Complies with the test for sterility (2.2.11). glacial acetic acid and 50 ml of methanol, add sufficient
Storage. Store protected from light and moisture at a methanol to produce 100.0 ml and dilute 1.0 ml of the resulting
temperature not exceeding 30°. If it is intended for use in the solution to 100.0 ml with methanol. Absorbance of the resulting
manufacture of parenteral preparations, the container should solution, measured within 30 minutes of preparation, at the
be sterile and sealed so as to exclude micro-organisms. maximum at about 305 nm, not less than 0.60. Use as the blank
a solution prepared in the same manner without the substance
Labelling. The label states whether or not the contents are under examination.
intended for use in the manufacture of parenteral preparations.
Heavy metals (2.3.13). 1.0 g complies with the limit test for
heavy metals, Method B (20 ppm).
Sulphated ash (2.3.18). Not more than 3.5 per cent.
Nystatin
Loss on drying (2.4.19). Not more than 5.0 per cent, determined
Nystatin is an antifungal substance produced by the growth on 1.0 g by drying in an oven at 60° at a pressure not exceeding
of certain strains of Streptomyces noursei or by any other 0.1 kPa for 3 hours.
means. It consists mainly of polyenes, the principal component
being nystatin A1. Assay. Protect the solution from light throughout the assay.
Nystatin has a potency of not less than 4400 Units per mg, Weigh accurately about 75 mg and dissolve in sufficient
calculated on the dried basis. dimethylformamide to produce 50.0 ml; dilute 10.0 ml of the
resulting solution to 200.0 ml with buffer solution No 4 (2.2.10).
Description. A yellow to slightly brown powder; odour,
characteristic; hygroscopic. Determine by the microbiological assay of antibiotics (2.2.10).

848
IP 2007 NYSTATIN TABLETS

Nystatin intended for oral administration complies with the Nystatin Pessaries
following additional requirement.
Nystatin Vaginal Tablets
Abnormal toxicity (2.2.1). Complies with the test for abnormal
toxicity, using a quantity containing not less than 600 Units Nystatin Pessaries contain not less than 90.0 per cent and not
suspended in not more than 0.5 ml of a 0.5 per cent w/v solution more than 130.0 per cent of the stated number of Units of
of acacia and injecting the suspension intraperitoneally. nystatin.

Storage. Store protected from light and moisture. Identification


Labelling. The label states the strength in terms of the number Extract a quantity of the powdered pessaries containing
of Units of Nystatin per mg. 300,000 Units with a mixture of 50 ml of methanol and 5 ml of
glacial acetic acid, add sufficient methanol to produce
100 ml and filter. Dilute 1 ml of the filtrate to 100 ml with
methanol. The resulting solution complies with the follwing
Nystatin Ointment test.
Disperse a quantity containing 25,000 Units in 10 ml of
Nystatin Ointment is a dispersion of Nystatin in microfine chloroform, add 40 ml of methanol and shake. Filter and dilute
powder in a suitable ointment basis. 1 ml of the filtrate to 25 ml with methanol.
Nystatin Ointment contains not less than 90.0 per cent and When examined in the range 230 nm to 360 nm (2.4.7), the
not more than 130.0 per cent of the stated number of Units of resulting solution shows absorption maxima at about 291 nm,
nystatin. 305 nm and 319 nm. The ratios of the absorbances at the maxima
at about 291 nm and about 319 nm to the absorbance at the
Identification maximum at about 305 nm are 0.61 to 0.73 and 0.83 to 0.96,
Disperse a quantity containing 25,000 Units in 10 ml of respectively. Use as the blank a solution prepared exactly in
chloroform, add 40 ml of methanol and shake. Filter and dilute the same manner without the substance under examination.
1 ml of the filtrate to 25 ml with methanol.
Tests
When examined in the range 230 nm to 360 nm (2.4.7), the
resulting solution shows absorption maxima at about 291 nm, Other Tests. Comply with the tests stated under Pessaries.
305 nm and 319 nm. The ratios of the absorbances at the maxima Loss on drying (2.4.19). Not more than 5 per cent, determined
at about 291 nm and about 319 nm to the absorbance at the on 1.0 g of the powdered pessaries by drying in an oven at 60°
maximum at about 305 nm are 0.61 to 0.73 and 0.83 to 0.96, at a pressure not exceeding 0.7 kPa for 3 hours.
respectively. Use as the blank a solution prepared exactly in
the same manner without the substance under examination. Assay. Protect the solution from light throughout the assay.
Weigh and powder 20 pessaries. Weigh accurately a quantity
Tests of the powder containing 200,000 Units and shake with 50.0 ml
Other tests. Complies with the tests stated under Ointments. of dimethylformamide for 1 hour. Centrifuge, dilute 10.0 ml of
the clear, supernatant liquid to 200.0 ml with buffer solution
Assay. Protect the solution from light throughout the assay. No 4 (2.2.10).
Weigh accurately a quantity containing 400,000 Units, disperse Determine by the microbiological assay of antibiotics (2.2.10).
in 20 ml of ether in a stoppered flask, add 70 ml of
Storage. Store protected from light and moisture.
dimethylformamide, shake for a few minutes, add 10 ml of
water, shake vigorously for a few minutes and add sufficient Labelling. The label states the strength in terms of the number
dimethylformamide to produce 100.0 ml. Mix well, filter and of Units of Nystatin.
dilute 10.0 ml of the filtrate to 100.0 ml with buffer solution No
4 (2.2.10).
Determine by the microbiological assay of antibiotics (2.2.10).
Nystatin Tablets
Storage. Store protected from moisture.
Nystatin Tablets contain not less than 90.0 per cent and not
Labelling. The label states the strength in terms of the number more than 130.0 per cent of the stated number of Units of
of Units of Nystatin per g. nystatin. The tablets are coated.

849
NYSTATIN TABLETS IP 2007

Identification tablets fail to disintegrate, wash them rapidly by immersion in


water and continue the test using phosphate buffer pH 6.8;
Extract a quantity of the powdered tablets containing the tablets then disintegrate within a further 30 minutes.
300,000 Units with a mixture of 50 ml of methanol and 5 ml of
glacial acetic acid, add sufficient methanol to produce Loss on drying (2.4.19). Not more than 5.0 per cent, determined
100 ml and filter. Dilute 1 ml of the filtrate to 100 ml with on 1.0 g of the powdered tablets by drying in an oven at 60° at
methanol. The resulting solution complies with the follwing a pressure not exceeding 0.7 kPa for 3 hours.
test. Other Tests. Comply with the tests stated under Tablets.
When examined in the range 230 nm to 360 nm (2.4.7), the Assay. Weigh and powder 20 tablets. Weigh accurately a
solution shows absorption maxima at about 291 nm, 305 nm quantity of the powder containing 200,000 Units and shake
and 319 nm. The ratios of the absorbances at the maxima at with 50.0 ml of dimethylformamide for 1 hour. Centrifuge, dilute
about 291 nm and about 319 nm to the absorbance at the 10.0 ml of the clear, supernatant liquid to 200.0 ml with buffer
maximum at about 305 nm are 0.61 to 0.73 and 0.83 to 0.96, solution No 4 (2.2.10).
respectively. Use as the blank a solution prepared exactly in
the same manner without the substance under examination. Determine by the microbiological assay of antibiotics (2.2.10).
Storage. Store protected from moisture.
Tests
Labelling. The label states the strength in terms of the number
Disintegration (2.5.1). 30 minutes, but using a 0.6 per cent of Units of Nystatin.
v/v solution of hydrochloric acid in place of water. If the

850
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

O
Oestradiol Benzoate ....
Oestradiol Injection ....
Ofloxacin ....
Ofloxacin Infusion ....
Ofloxacin Opthalmic Solution ....
Ofloxacin Tablets ....
Olanzapine ....
Olanzapine Tablets ....
Oleic Acid ....
Omeprazole ....
Omeprazole Capsules ....
Oral Rehydration Salts ....
Ormeloxifen Hydrochloride ....
Ormeloxifen Hydrochloride Tablets ....
Orphenadrine Citrate ....
Orphenadrine Hydrochloride ....
Orphenadrine Tablets ....
Oseltamivir Phosphate ....
Oseltamivir Capsules ....
Oseltamivir Oral Suspension ....
Oxazepam ....
Oxazepam Tablets ....
Oxprenolol Hydrochloride ....
Oxprenolol Tablets ....
Oxygen ....
Oxygen 93 Per Cent ....
Oxyphenbutazone ....
Oxyphenbutazone Tablets ....
Oxytetracycline Dihydrate ....
Oxytetracycline Injection ....

851
MONOGRAPHS INDIAN PHARMACOPOEIA 2007 2007

Oxytetracycline Hydrochloride ....


Oxytetracycline Capsules ....
Oxytetracycline Eye Ointment ....
Oxytetracycline Hydrochloride Injection ....
Oxytocin ....
Oxytocin Injection ....
Oxytocin Nasal Solution ....

852
IP 2007 OESTRADIOL INJECTION

Oestradiol Benzoate Test solution (b). Dissolve 0.1 g of the substance under
examination in 100 ml of the same solvent mixture.

OH Reference solution (a). A 0.02 per cent w/v solution of the


H3C
substance under examination in the same solvent mixture.
H Reference solution (b). A 0.1 per cent w/v solution of
O oestradiol benzoate RS in the same solvent mixture.
H H
Apply to the plate 5 µl of each solution. After development,
O
dry the plate in air until the odour of the solvent is no longer
detectable, heat at 110º for 10 minutes, spray the plate while
hot with ethanolic sulphuric acid (20 per cent), heat again at
C25H28O3 Mol. Wt. 376.5 110º for 10 minutes and examine in ultraviolet light at 365 nm.
Any secondary spot in the chromatogram obtained with test
Oestradiol Benzoate is 17β-hydroxyestra-1,3,5(10)-trien-3-yl
solution (a) is not more intense than the spot in the
benzoate.
chromatogram obtained with reference solution (a).
Oestradiol Benzoate contains not less than 97.0 per cent and
Sulphated ash (2.3.18). Not more than 0.2 per cent, determined
not more than 103.0 per cent of C25H28O3, calculated on the
on 0.5 g.
dried basis.
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
Description. Colourless crystals or a white, crystalline powder;
on 0.5 g by drying in an oven at 105º for 3 hours.
odourless.
Assay. Weigh accurately about 25 mg, dissolve in sufficient
Identification ethanol (95 per cent) to produce 250.0 ml. Dilute 10.0 ml to
100.0 ml with ethanol (95 per cent) and measure the
Test A may be omitted if tests B and C are carried out. Test C
absorbance of the resulting solution at the maximum at about
may be omitted if tests A and B are carried out.
231 nm (2.4.7). Calculate the content of C25H28O3 taking 500 as
A. Determine by infrared absorption spectrophotometry (2.4.6). the specific absorbance at 231 nm.
Compare the spectrum with that obtained with oestradiol Storage. Store protected from light and moisture.
benzoate RS or with the reference spectrum of oestradiol
benzoate.
B. In the test for Related substances, the principal spot in the
chromatogram obtained with test solution (b) corresponds to Oestradiol Injection
that in the chromatogram obtained with reference solution (b)
when examined in daylight and in ultraviolet light at 365 nm. Oestradiol Benzoate Injection
C. To about 1 mg add 0.5 ml of a 5 per cent w/v solution of Oestradiol Injection is a sterile solution of Oestradiol Benzoate
ammonium molybdate in sulphuric acid; a yellowish green in Ethyl Oleate or other suitable ester, in a suitable fixed oil or
colour develops which exhibits an intense green fluorescence in any mixture of these. It may contain suitable alcohols.
when examined in ultraviolet light at 365 nm. Add 1 ml of Oestradiol Benzoate Injection contains not less than 90.0 per
sulphuric acid and 9 ml of water; the solution becomes pink cent and not more than 110.0 per cent of the stated amount of
with a yellowish fluorescence. oestradiol benzoate, C25H28O3.
Tests Identification
Specific optical rotation (2.4.22). +57.0º to +63.0º, determined Determine by thin-layer chromatography (2.4.17), coating the
in a 1.0 per cent w/v solution in dioxan. plate with silica gel G.
Related substances. Determine by thin-layer chromatography Mobile phase. A mixture of 80 volumes of toluene and
(2.4.17), coating the plate with silica gel G. 20 volumes of ethyl acetate.
Mobile phase. A mixture of 90 volumes of toluene and Test solution. Add 10 ml of 2,2,4-trimethylpentane to a volume
10 volumes of ethanol (95 per cent). of the injection containing 2 mg of Oestradiol Benzoate and
Test solution (a). Dissolve 0.2 g of the substance under extract with three quantities, each of 10 ml, of ethanol (70 per
examination in 10 ml of a mixture of 90 volumes of chloroform cent). Wash the combined extracts with 15 ml of 2,2,4-
and 10 volumes of methanol. trimethylpentane, evaporate the ethanolic extract to dryness

853
OFLOXACIN IP 2007

using a rotary evaporator and dissolve the residue in 2 ml of Ofloxacin


chloroform.
Reference solution. A 0.1 per cent w/v solution of oestradiol H 3C CH3
N O
benzoate RS in chloroform.
N N
Apply to the plate 5 µl of each solution. After development,
dry the plate in air, spray with ethanolic sulphuric acid
(20 per cent), heat at 105º for 10 minutes and examine in F COOH
ultraviolet light at 365 nm. The principal spot in the O
chromatogram obtained with the test solution corresponds to
C18H20FN3O4 Mol. Wt. 361.4
that in the chromatogram obtained with the reference solution.
Ofloxacin is (RS)-9-fluoro-3-methyl-10-(4-methylpiperazin-1-
Tests yl)-7-oxo-2,3-dihydro-7H-pyrido[1,2,3,-de]-1,4-benzoazeine-
6-carboxylic acid
Other Tests. Complies with the tests stated under Parenteral
Preparations (Injections). Ofloxacin contains not less than 98.5 per cent and not more
than of 101.5 per cent of C18H20FN3O4, calculated on the dried
Assay. Determine by liquid chromatography (2.4.14).
basis.
Test solution (a). Dilute an accurately measured quantity of
Description. A pale yellow or bright yellow, crystalline powder.
the injection containing about 1 mg of Oestradiol Benzoate to
10.0 ml with a mixture of 90 volumes of cyclohexane and Identification
10 volumes of dioxan.
Determine by infrared absorption spectrophotometry (2.4.6).
Test solution (b). Add 1 ml of a solution prepared by dissolving
Compare the spectrum with that obtained with ofloxacin RS.
15 mg of 4-hydroxybenzaldehyde (internal standard) in
10.0 ml of dioxan, adding sufficient cyclohexane to produce Tests
100.0 ml (solution A), to an accurately measured quantity of
the injection containing about 1 mg of Oestradiol Benzoate Light absorption. Absorbance at 440 nm (2.4.7), of 0.5 per cent
and dilute to 10.0 ml with sufficient of a mixture of 90 volumes w/v solution in 0.1 M hydrochloric acid is not more than 0.25.
of cyclohexane and 10 volumes of dioxan. Related substances. Determine by liquid chromatography
Reference solution. Add 10 ml of solution A to 10 mg of (2.4.14).
oestradiol benzoate RS, accurately weighed, and dilute to Test solution. Dissolve 10 mg of the substance under
100.0 ml with the same solvent mixture. examination in 10 ml of methanol.
Chromatographic system Reference solution (a). A 0.1 per cent w/v solution of
– a stainless steel column 30 cm x 4 mm, packed with porous ofloxacin RS in methanol.
silica particles (10 µm), Reference solution (b). Dilute 1 ml of reference solution (a) to
– mobile phase: 90 volumes of cyclohexane and 100 ml with methanol.
10 volumes of dioxan,
– flow rate. 2 ml per minute, Chromatographic system
– spectrophotometer set at 254 nm, – a stainless steel column 10 cm x 4.6 mm packed with
– a 20 µl loop injector. octadecylsilyl silica gel for chromatography (5 µm),
– mobile phase: a mixture of 10 volumes of acetonitrile
Inject test solutions (a), (b) and the reference solution. The and 90 volumes of phosphate buffer pH 2.4 prepared by
assay is not valid unless the resolution between the peaks dissolving 27.2 g of monobasic potassium phosphate
due to benzyl alcohol (if present) and oestradiol benzoate and in 1000 ml of water, adjust the pH to 2.4 with
between the peaks due to oestradiol benzoate and the internal orthophosphoric acid,
standard is more than 1.5. – flow rate. 2 ml per minute,
Calculate the content of C25H28O3 in the injection. – spectrophotometer set at 294 nm,
– a 10 µl loop injector.
Storage. Store protected from light.
Inject reference solution (b). The test is not valid unless the
Labelling. The label states (1) the nature and composition of
column efficiency in not less than 1400 theoretical plates.
the solvent; (2) that it is meant for intramuscular injection
only; (3) that any solid matter that may have separated on Inject the test solution and reference solution (b). Run the
standing should be redissolved by warming before use. chromatogram three times of the principal peak. In the

854
IP 2007 OFLOXACIN OPHTHALMIC SOLUTION

chromatogram obtained with the test solution, the area of any – mobile phase: a mixture of 80 volumes of buffer solution
secondary peak is not more than 0.5 times the area of the peak prepared by dissolving 6.8 g of potassium dihydrogen
in the chromatogram obtained with reference solution (b) (0.5 orthophosphate and 0.47 g sodium 1-hexane
per cent) and the sum of areas of all the secondary peaks is sulphonate in 1000 ml of water, add 1 ml of triethylamine
not more than the area of the peak in the chromatogram and adjust the pH to 3.0 with orthophosphoric acid
obtained with the reference solution (b) (1.0 per cent). and 20 volumes of acetonitrile,
Heavy Metals (2.3.13). 2.0 g complies with the limit test for – flow rate. 1 ml per minute,
heavy metals, Method C (10 ppm). – spectrophotometer set at 294 nm,
– a 20 µl loop injector.
Sulphated ash (2.3.18). Not more than 0.1 per cent.
Inject the reference solution. The test is not valid unless the
Loss on drying (2.4.19). Not more than 0.2 per cent, determined tailing factor is not more than 2.0 and the relative standard
on 1.0 g by drying in an oven at 105° for 4 hours. deviation for replicate injections is not more than 2.0 per cent.
Assay. Weigh accurately about 0.3 g, dissolve in 100 ml Inject the test solution and the reference solution.
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
Calculate the content of C18H20FN3O4 in infusion.
acid, determining the end-point potentiometrically (2.4.25).
Carry out a blank titration.
1 ml 0.1 M perchloric acid is equivalent of 0.03614 g of
C18H20FN3O4.
Ofloxacin Ophthalmic Solution
Ofloxacin Ophthalmic Solution is a sterile aqueous solution of
Storage. Store protected from light and moisture.
Ofloxacin.
Ofloxacin Ophthalmic Solution contain not less than 90.0 per
cent and more than 110.0 per cent of the stated amount of
Ofloxacin Infusion ofloxacin, C18H20FN3O4.
Ofloxacin Infusion contain Ofloxacin in water for injection. Identification
Ofloxacin Infusion contain not less than 90.0 per cent and not In the Assay, the principal peak in the chromatogram obtained
more than 120.0 per cent of the stated amount of ofloxacin, with the test solution corresponds to the peak in the
C18H20FN3O4. chromatogram obtained with the reference solution.
Identification Tests
In the Assay, the principal peak in the chromatogram obtained pH (2.4.24). 6.0 to 7.2.
with the test solution corresponds to the peak in the Sterility (2.2.11). Comply with the test for sterility.
chromatogram obtained with the reference solution.
Assay. Determine by liquid chromatography (2.4.14).
Tests Solvent mixture. Phosphate buffer pH 7.25 prepared by
dissolving 3.56 g of disodium hydrogen phosphate in 1000 ml
pH (2.4.24). 3.8 to 5.8
water, adjusted pH to 7.25 using orthophosphoric acid or
Other tests. Complies with the tests stated under Parenteral sodium hydroxide solution.
Preparation (Infusions).
Test solution. Measure accurately a volume of Opthalmic
Assay. Determine by liquid chromatography (2.4.14). Solution containing 30 mg of Ofloxacin in 100.0 ml of solvent
NOTE —Protect the solutions from the light. mixture. Dilute 1.0 ml of the solution to 10.0 with solvent
mixture.
Test solution. Measure accurately a volume containing 50 mg
Reference solution. A 0.003 per cent w/v solution of ofloxacin
of Ofloxacin in 100.0 ml with mobile phase. Dilute 5.0 ml of the
RS in solvent mixture.
solution to 50.0 ml with mobile phase.
Chromatographic system
Reference solution. A 0.005 per cent w/v solution of ofloxacin
– a stainless steel column 15 cm x 4.6 mm packed with
RS in mobile phase.
octadecylsilane bonded to porous silica (5 µm) (such as
Chromatographic system TSK GEL),
– a stainless steel column 25 cm x 4.6 mm packed with – mobile phase: a mixture of 20 volumes of acetonitrile,
octadecylsilane bonded to porous silica (5 µm), 80 volumes of phosphate buffer pH 7.25 prepared by

855
OFLOXACIN TABLETS IP 2007

dissolving 2.54 g of tetrabutyl ammonium hydrogen Reference solution (b). Dilute 1 ml of reference solution (a) to
sulphate and 3.56 g of disodium hydrogen phosphate 100 ml with methanol.
in 1000 ml water, Chromatographic system
– flow rate. 1.5 ml per minute, – a stainless steel column 10 cm x 4.6 mm packed with
– spectrophotometer set at 254 nm, octadecylsilyl silica gel for chromatography (5 µm),
– a 20 µl loop injector. – mobile phase: a mixture of 10 volumes of acetonitrile
Inject the reference solution. The test is not valid unless the and 90 volumes of phosphate buffer pH 2.4 prepared by
relative standard deviation for replicate injections is not more dissolving 27.2 g of monobasic potassium phosphate
than 2.0 per cent. in 1000 ml of water, adjust the pH to 2.4 with
orthophosphoric acid,
Inject the test solution and the reference solution.
– flow rate. 2 ml per minute,
Calculate the content of C18H20FN3O4. – spectrophotometer set at 294 nm,
Storage. Store protected from light. – a 10 µl loop injector.
Inject reference solution (a). The test is not valid unless the
column efficiency in not less than 1400 theoretical plates.
Ofloxacin Tablets Inject the test solution and reference solution (b). Run the
chromatogram three times of the principal peak. In the
Ofloxacin Tablets contain Ofloxacin. chromatogram obtained with the test solution, the area of any
Ofloxacin Tablets contain not less than 90.0 per cent and not secondary peak is not more than the area of the peak in the
more than 110.0 per cent of the stated amount of ofloxacin, chromatogram obtained with reference solution (b) (1.0 per
C18H20FN3O4. cent) and the sum of areas of all the secondary peaks is not
more than twice the area of the peak in the chromatogram
Identification obtained with the reference solution (b) (2.0 per cent).

In the Assay, the principal peak in the chromatogram obtained Other tests. Comply with the tests stated under the Tablets.
with the test solution corresponds to the peak in the Assay. Determine by liquid chromatography (2.4.14).
chromatogram obtained with the reference solution (a).
Test solution. Weigh and powder 20 tablets. Weigh accurately
Tests a quantity of powdered tablet containing 25 mg of Ofloxacin,
disperse in 60 ml of methanol and dilute to 100 ml with methanol
Dissolution (2.5.2). and filter.
Apparatus. No 1 Reference solution. A 0.025 per cent w/v solution of ofloxacin
Medium. 900 ml of 0.1 M hydrochloride acid. RS in methanol.
Speed and time. 50 rpm for 30 minutes. Chromatographic system
Withdraw a suitable volume of the medium and filter. Measure – a stainless steel column 15 cm x 4.6 mm packed with
the absorbance of the filtrate, suitably diluted with the medium octadecylsilane bonded to porous silica (5 µm),
if necessary, at the maximum at about 294 nm (2.4.7). Calculate – mobile phase: a mixture of 92 volumes of buffer solution
the content of C18H20FN3O4 in the medium from the absorbance prepared by dissolving 27.2 g of potassium dihydrogen
obtained from a solution of known concentration of phosphate in 1000 ml of water and adjust the pH to 2.4
ofloxacin RS in the same medium. with orthophosphoric acid and 8 volumes of
acetonitrile,
D. Not less than 75 per cent of the stated amount of
– flow rate. 2 ml per minute,
C18H20FN3O4.
– spectrophotometer set at 294 nm,
Related substances. Determine by liquid chromatography – a 10 µl loop injector.
(2.4.14).
Inject the reference solution. The test is not valid unless the
Test solution. Weigh and powder 20 tablets. Weigh accurately relative standard deviation for replicate injections is not more
a quantity of powdered tablet containing 100 mg of Ofloxacin, than 2.0 per cent.
disperse in 60 ml of methanol and dilute to 100 ml with methanol
Inject the test solution and the reference solution.
and filter.
Calculate the content of C18H20FN3O4.
Reference solution (a). A 0.1 per cent w/v solution of
ofloxacin RS in methanol. Storage. Store protected from light and moisture.

856
IP 2007 OLANZAPINE TABLETS

Olanzapine – spectrophotometer set at 230 nm,


– a 10 µl loop injector.
Time Mobile phase A Mobile phase B
CH3 (in min.) ( per cent v/v) ( per cent v/v)
N 0 100 0
20 63 37
N
N 30 45 55
32 100 0
N CH3 38 100 0
S
H Inject reference solution (b). The test is not valid unless the
tailing factor is not more than 2.0 and the column efficiency in
C17H20N4S Mol. Wt. 312.4 not less than 2000 theoretical plates.

Olanzapine is 2-methyl-4-(4-methyl-1-piperazinyl)-10H- Inject the test solution and reference solution (b). Run the
thieno[2,3-b][1,5]benzodiazepine chromatogram for three times, the principal peak due to
olanzapine. In the chromatogram obtained with the test
Olanzapine contains not less than 98.0 per cent and not more solution, the area of any secondary peak is not more than 0.5
than 102.0 per cent of C17H20N4S, calculated on the anhydrous times the area of the peak in the chromatogram obtained with
basis. reference solution (b) (0.5 per cent) and the sum of areas of all
Description. A yellow crystalline powder. the secondary peaks is not more than twice the area of the
peak in the chromatogram obtained with the reference solution
Identification (b) (2.0 per cent).
Determine by infrared absorption spectrophotometry (2.4.6). Heavy metals (2.3.13). 1.0 g complies with the limit test for
Compare the spectrum with that obtained with olanzapine RS heavy metals, Method B (20 ppm).
or with the reference spectrum of olanzapine. Sulphated ash (2.3.18). Not more than 0.1 per cent.
Tests Water (2.3.43). Not more than 1.0 per cent, determined on
1.0 g.
Related substances. Determine by liquid chromatography
(2.4.14). Assay. Weigh accurately about 0.2 g, dissolve in 40 ml of
glacial acetic acid. Titrate with 0.1 M perchloric acid,
Solvent mixture. 40 volumes of water and 60 volumes of determining the end-point potentiometrically (2.4.25). Carry
acetonitrile. out a blank titration.
Test solution. Dissolve 50 mg of the substance under
1 ml of 0.1 M perchloric acid is equivalent to 0.01562 g of
examination in 25 ml of solvent mixture.
C17H20N4S.
Reference solution (a). A 0.2 per cent w/v solution of
Storage. Store protected from light and moisture, at a
olanzapine RS in solvent mixture.
temperature not exceeding 30º.
Reference solution (b). Dilute 1 ml of reference solution (a) to
100 ml with solvent mixture.
Chromatographic system
– a stainless steel column 25 cm x 4.6 mm packed with Olanzapine Tablets
octadecylsilane bonded to porous silica (5 µm),
Olanzapine Tablets contain not less than 90.0 per cent and not
– column temperature 40º,
more than 110.0 per cent of the stated amount of olanzapine,
– mobile phase: A. a mixture of 80 volumes of buffer
C17H20N4S.
solution pH 6.8 prepared by dissolving 4.825 g of
sodium dihydrogen orthophosphate monohydrate in
Identification
1000 ml of water, adjust pH to 6.8 with 10 per cent w/v of
sodium hydroxide and 20 volumes of acetonitrile, In the Assay, the principal peak in the chromatogram obtained
B. acetonitrile, with the test solution corresponds to the peak in the
– flow rate. 1.2 ml per minute, chromatogram obtained with the reference solution.

857
OLEIC ACID IP 2007

Tests less than 2500 theoretical plates. The relative standard


deviation for replicate injections is not more than 2 per cent.
Dissolution (2.5.2).
Inject the test solution and the reference solution.
Apparatus. No 1
Medium. 900 ml of 0.01 M hydrochloric acid. Calculate the content of C17H20N4S.
Speed and time. 50 rpm for 45 minutes. Storage. Store protected from light and moisture, at a
Withdraw a suitable volume of the medium and filter. Determine temperature not exceeding 25º.
by liquid chromatography (2.4.14).
NOTE – Protect all the solutions from light.
Test solution. Use the filtrate, if necessary dilute with Oleic Acid
dissolution medium.
Oleic Acid consists mainly of (Z)-octadec-9-enoic acid,
Reference solution. Weigh 16 mg of olanzapine RS, dissolve
C18H34O2, together with varying amounts of saturated and
in about 2.5 ml of acetonitrile and dilute to 25 ml with 0.01 M
other unsaturated fatty acids and is obtained by the hydrolysis
hydrochloroc acid.Dilute suitabely to get 0.0016 per cent w/v
of fats or fixed oils and separation of the liquid acids by
in dissolution medium.
expression or other suitable means. It may contain a suitable
Chromatographic system as described under Assay, using 50 antioxidant.
µl loop injector.
Description. A clear, yellowish to pale brown, oily liquid; odour,
Inject the reference solution . The test is not valid unless the characteristic. On exposure to air it darkens in colour and the
tailing factor is not more than 2.0. The column efficiency in odour becomes more pronounced.
not less than 2500 theoretical plates.
Inject the test solution and the reference solution. Identification
Calculate the content of C17H20N4S. A.To 1 ml add 1 ml of ethanol (95 per cent); the solution is
D. Not less than 70 per cent of the stated amount of C17H20N4S. clear. It turns orange or red on addition of 0.1 ml of methyl
orange solution.
Other tests. Comply with the tests stated under Tablets.
B.Take a mixture of 1 ml of nitric acid and 1 ml of water in a
Assay. Determine by liquid chromatography (2.4.14). test-tube with an internal diameter of about 12.5 mm and add
Test solution. Take 10 intact tablets to a suitable volumetric 1 ml of the substance under examination on the surface of the
flask, disperse in acetonitrile and dilute with 0.01 M mixture. Introduce 0.5 g of copper turnings into the lower
hydrochloroc acid to get final concentration of 0.01 per cent layer and allow to stand under a hood for 4 hours; the upper
w/v of Olanzapine. layer solidifies.
Reference solution. Weigh 10 mg of olanzapine RS, dissolve C.Complies with the test for Iodine value (2.3.28).
in about 25 ml of acetonitrile and dilute to 100 ml with 0.01 M
hydrochloroc acid. Tests
Chromatographic system Weight per ml (2.4.29). 0.889 g to 0.895 g.
– a stainless steel column 25 cm x 4.6 mm packed with
packed with octadecylsilyl silica gel for chromatography Peroxide value (2.3.35). Not more than 10.0.
(5 µm), Acid value (2.3.23). 195 to 202, determined on 0.5 g.
– mobile phase: a mixture of 70 volumes of buffer solution
Iodine value (2.3.28). 85 to 95, determined by Method A.
prepared by dissolving 3 g of ammonium dihydrogen
orthophosphate in 900 ml water, add 2 ml of Water-soluble acids. Shake 5 ml with 5 ml of water for
triethylamine and dilute to 1000 ml with water. Adjust 2 minutes, allow the liquids to separate and filter the aqueous
pH to 2.5 with orthophosphoric acid and 30 volumes layer through paper moistened with water. To the filtrate add
of methanol, 0.05 ml of methyl orange solution; the liquid does not become
– flow rate. 1 ml per minute, red.
– spectrophotometer set at 220 nm,
Neutral fats and mineral oils. Boil 1 ml with 5 ml of 0.5 M
– a 10 µl loop injector.
sodium carbonate and 25 ml of water in a large flask. The
Inject the reference solution. The test is not valid unless the solution, while still hot, is not more opalescent than
tailing factor is not more than 2.0, the column efficiency in not opalescence standard OS2 (2.4.1).

858
IP 2007 OMEPRAZOLE

Congealing point. Dry about 10 ml by heating to 110º with Tests


frequent stirring, transfer to a test-tube about 20 mm in diameter,
cool and when at 15º immerse the tube in a suitable water-bath Appearance of solution. A 2.0 per cent w/v solution in
so that the cooling takes place at the rate of 2º per minute. Stir dichloromethane is clear (2.4.1).
the sample with a thermometer; it does not become cloudy Light absorption (2.4.7). Absorbances of a freshly prepared
until the temperature has fallen to 10º. On further cooling it 2.0 per cent w/v solution in dichloromethane at 400 nm,
congeals to a white solid or semi-solid mass at about 4º. 500 nm and 600 nm are not more than 0.25, 0.10 and 0.10
Sulphated ash (2.3.18). Not more than 0.1 per cent. respectively.
Related substances. A. Determine by thin-layer chromato-
Storage. Store protected from light and moisture in a refrigerator
graphy (2.4.17), coating the plate with silica gel GF254.
(8º to 15º).
Mobile phase. A mixture of 50 volumes of benzene, 30 volumes
Labelling. The label states (1) where applicable, that it is used
of ethyl acetate and 10 volumes of methanol.
for external use only; (2) the name and concentration of any
added antioxidant. Test solution. Dissolve 0.4 g of the substance under
examination in 100 ml of ethanol.
Reference solution (a). A 0.4 per cent w/v solution of
omeprazole RS in ethanol.
Omeprazole Reference solution (b). A 0.004 per cent w/v solution of
omeprazole RS in ethanol.
H CH3 Reference solution (c). A 0.002 per cent w/v solution of
N O N
omeprazole RS in ethanol.
S
N OCH3 Apply to the plate 10 µl of each solution. After development,
H3CO
CH3 dry the plate in air and examine in ultraviolet light at 254 nm
immediately. Any secondary spot in the chromatogram
C17H19N3O3S Mol. Wt. 345.4 obtained with the test solution is not more intense than the
spot in the chromatogram obtained with reference solution (c)
Omeprazole is 5-methoxy-2-[[(4-methoxy-3,5- and the total intensity of all such spots in the chromatogram
dimethylpyridin-2-yl)methyl]sulfinyl] -1H-benzimidazole. obtained with the test solution is not more than the intensity
Omeprazole contains not less than 99.0 per cent and not more of the spot obtained with reference solution (b).
than 101.0 per cent of C17H19N3O3S, calculated on the dried B. In the Assay, the sum of the areas of all the secondary
basis. peaks is not greater than 1.5 per cent of the total area of all
NOTE — Perform the tests and assay in subdued light and peaks.
use low-actinic glassware. Heavy metals (2.3.13). The residue obtained in the test for
Description. A white or almost white powder. Sulphated ash, complies with the limit test for heavy metals,
Method B (20 ppm).
Identification Sulphated ash (2.3.18). Not more than 0.2 per cent.
Test A may be omitted if tests B and C are carried out. Tests B Loss on drying (2.4.19). Not more than 0.2 per cent, determined
and C may be omitted if test A is carried out. on 1.0 g by drying in an oven at 60º at a pressure not exceeding
0.7 kPa for 4 hours.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with omeprazole Assay. Determine by liquid chromatography (2.4.14).
RS or with the reference spectrum of omeprazole. Test solution. A 0.005 per cent w/v solution of the substance
B. When examined in the range 230 nm to 360 nm (2.4.7), a under examination in the mobile phase.
0.002 per cent w/v solution in 0.1 M sodium hydroxide shows Reference solution. A 0.005 per cent w/v solution of
absorption maxima at about 276 nm and 305 nm; the ratio of omeprazole RS in the mobile phase.
the absorbance at about 305 nm to that at about 276 nm, 1.6 to
Chromatographic system
1.8.
– a stainless steel column 30 cm x 3.9 mm, packed with
C. In the test for Related substances, the principal spot in the octadecylsilane bonded to porous silica (5 µm),
chromatogram obtained with the test solution corresponds to – mobile phase: a mixture of 65 volumes of phosphate
that in the chromatogram obtained with reference solution (a). buffer pH 7.4 and 35 volumes of acetonitrile,

859
OMEPRAZOLE CAPSULES IP 2007

– flow rate. 1 ml per minute, for 2 hours, drain the solution slowly without losing any
– spectrophotometer set at 302 nm, granules. Transfer them quantitatively to a 100-ml volumetric
– a 20 µl loop injector. flask, add 20 ml of 0.1 M sodium hydroxide and mix with the
Inject the reference solution. Repeat the procedure at least aid of ultrasound. Dilute to volume with 0.1 M sodium
five times and measure the peak responses of the peak due to hydroxide, centrifuge about 15 ml for 5 minutes and dilute
omeprazole. The relative standard deviation of the replicate 5.0 ml of the clear supernatant liquid to 50.0 ml with the mobile
injections is not more than 2.0 per cent. phase. Using the resulting solution as the test solution, carry
out the determination as described in the Assay. Calculate the
Calculate the content of C17H19N3O3S. content of C17H19N3O3S in the supernatant liquid. Calculate
Storage. Store protected from light and moisture in a refrigerator the percentage of omeprazole released in the acid medium by
(2º to 8º). subtracting the content of C17H19N3O3S in the test solution
from the total content of omeprazole determined in the Assay.
NOTE — A combination of elevated temperatures (37º-50º)
and high humidity degrades Omeprazole. It rapidly degrades Not more than 15 per cent of the stated amount of C17H19N3O3S
under acidic conditions. is dissolved in 2 hours.
B. Apparatus. No 1
Medium. 900 ml of phosphate buffer pH 6.8,
Omeprazole Capsules Speed and time. 100 rpm and 45 minutes.
Omeprazole Capsules contain enteric-coated granules of Tap the granules from a capsule slightly with a glass rod to
Omeprazole. make them settle to the bottom. Rotate the paddle at 100 rpm
for 45 minutes and filter the solution. Using the filtered medium
Omeprazole Capsules contain not less than 90.0 per cent and as the test solution, carry out the determination as described
not more than 110.0 per cent of the stated amount of in the Assay. Calculate the content of C17H19N3O3S in the
omeprazole, C17H19N3O3S. medium.
NOTE — Perform the tests and assay in subdued light and D. Not less than 70 per cent of the stated amount of
use low-actinic glassware. C17H19N3O3S.
Identification Other Tests. Comply with the tests stated under Capsules.
Loss on drying (2.4.19). Not more than 3.0 per cent, determined
A.To a quantity of the contents of the capsules containing
on 0.5 g of the contents of the capsules by drying in an oven
50 mg of Omeprazole in a 100-ml volumetric flask add about
at 60º at a pressure not exceeding 0.7 kPa for 4 hours.
70 ml of 0.1 M sodium hydroxide. Mix in an ultrasonic bath for
about 5 minutes and heat on a water-bath for 10 minutes. Assay. Determine by liquid chromatography (2.4.14).
Cool, make up to volume with 0.1 M sodium hydroxide and Test solution. Mix the contents of 20 capsules. Weigh and
filter. Dilute 2 ml of the filtrate to 100 ml with 0.1 M sodium transfer the granules containing about 20 mg of Omeprazole
hydroxide. to a 100-ml volumetric flask, add 20 ml of 0.1 M sodium
When examined in the range 230 nm to 360 nm (2.4.7), the hydroxide, mix with the aid of ultrasound and dilute to volume
resulting solution shows absorption maxima at about 276 nm with 0.1 M sodium hydroxide. Centrifuge for 5 minutes and
and 305 nm. dilute 5.0 ml of the clear supernatant liquid to 50.0 ml with the
mobile phase.
B. In the Assay, the principal peak in the chromatogram
obtained with the test solution corresponds to the peak due Reference solution. Take 20 mg of omeprazole RS in a dry,
to omeprazole in the chromatogram obtained with the reference stoppered test-tube, add 20.0 ml of 0.1 M sodium hydroxide,
solution. shake vigorously for 5 minutes and dilute 1.0 ml of the solution
with the mobile phase to produce 50.0 ml.
Tests Chromatographic system
Dissolution (2.5.2). – a stainless steel column 30 cm x 3.9 mm, packed with
octadecylsilane bonded to porous silica (5 µm),
A. Apparatus. No 1
– mobile phase: a mixture of 65 volumes of phosphate
Medium. 900 ml of 0.1 M hydrochloric acid,
buffer pH 7.4 and 35 volumes of acetonitrile,
Speed and time. 100 rpm and 2 hours. – flow rate. 1 ml per minute,
Tap the granules from a capsule slightly with a glass rod to – spectrophotometer set at 302 nm,
make them settle to the bottom. Rotate the paddle at 100 rpm – a 20 µl loop injector.

860
IP 2007 ORAL REHYDRATION SALTS

Inject the reference solution. Repeat the procedure at least Oral Rehydration Salts contain not less than 90.0 per cent and
five times and measure the peak responses of the peak due to not more than 110.0 per cent of the stated amount of Dextrose
omeprazole. The relative standard deviation of the replicate (anhydrous) or Dextrose Monohydrate (as appropriate) and
injections is not more than 2.0 per cent. of the requisite amounts of sodium, Na, potassium, K, chloride,
Calculate the content of C17H19N3O3S in the capsules. Cl, and citrate, C6H5O7, calculated from the stated amounts of
the relevant constituents.
Storage. Store protected from light and moisture.
Description. A white to creamy-white, amorphous or
crystalline powder; odourless.

Oral Rehydration Salts Identification


ORS Powder A. When heated, melting and charring occurs and an odour of
burnt sugar is produced.
Oral Rehydration Salts are dry, homogeneously mixed powders
containing Dextrose, Sodium Chloride, Potassium Chloride B. Add a few drops of the solution prepared as directed in the
and either Sodium Bicarbonate or Sodium Citrate for use in label to 5 ml of potassium cupri-tartrate solution; a copious
oral rehydration therapy after being dissolved in the requisite red precipitate is produced on boiling.
amount of water. C. Gives reaction A of sodium salts, reaction A of potassium
They may contain suitable flavouring agents and, where salts and reaction A of chlorides (2.3.1).
necessary, suitable flow agents in the minimum quantity D. A quantity containing about 50 mg of citric acid gives the
required to achieve a satisfactory product but may not contain reactions of citrates (2.3.1).
artificial sweetening agents like mono- and/or poly-
saccharides. If saccharin/saccharin sodium or aspartame is Tests
used in preparations meant for paediatric use, the concentration Uniformity of weight. Comply with the test for contents of
of saccharin should be such that its daily intake is not more packaged dosage forms (2.5.6).
than 5 mg/kg of body weight and that of aspartame should be
such that its daily intake is not more than 40 mg/kg of body Seal test (only for sachets). Loosely bundle 10 sachets with a
weight. rubber band and submerge the bundle under water in a vacuum
desiccator maintained at a pressure not exceeding 18 kPa for
Strength. A formulation of reduced osmolarity (given below) one minute. Examine the bundle for any fine stream of bubbles.
recommended by the World Health Organization (WHO) for Re-establish normal pressure and open the bundle. No
the Diarrhoeal Diseases Control Programme, and of the United penetration of water is observed in any sachet.
Nations Children’s Fund (UNICEF).
Other Tests. Comply with the tests stated under Oral Powders.
Composition of the formulation in terms of the amount, in g, to
be dissolved in sufficient water to produce 1000 ml. Assay. Carry out the following assays on the well-mixed
contents of an individual sachet or on a suitable sample from
Sodium Chloride 2.6 the well-mixed contents of a bulk container. Where the amount
Dextrose (anhydrous) 13.5 in an individual sachet is insufficient to carry out all the assays,
or take a separate sachet for the Assay for citrate and for the
Dextrose Monohydrate 14.85 Assay for dextrose. For the Assays for total sodium, for
Potassium Chloride 1.5 potassium and for total chloride weigh accurately about 8.0 g
Sodium Citrate 2.9 and dissolve in sufficient water to produce 500.0 ml (solution
A).
The molar concentrations of sodium, potassium, chloride and
citrate ions in terms of millimoles per litre are given below: For total sodium — Dilute a suitable volume of solution A
mmol/l with a sufficient volume of a solution of strontium chloride
such that the final solution contains a 1500- to 2000-fold excess
Sodium 75
of strontium ions and determine by Method A for atomic
Potassium 20 absorption spectrophotometry (2.4.2), measuring at 589 nm
Chloride 65 and using sodium solution AAS, suitably diluted with the
Citrate 10 strontium chloride solution, for the standard solutions or,
Dextrose 75 alternately by Method A for flame photometry (2.4.4).
The total osmolar concentration of the solution in terms of 1 g of Sodium Chloride, and of Sodium Citrate is equivalent to
mOsmol per litre is 245. 0.3934 and 0.2345 g of Na respectively.

861
ORMELOXIFEN HYDROCHLORIDE IP 2007

For potassium — Dilute a suitable volume of solution A with Ormeloxifen Hydrochloride


a sufficient volume of solution of strontium chloride such
that the final solution contains a 1500- to 2000-fold excess of Centchroman Hydrochloride; Centchroman
strontium ions and determine by Method A for atomic
absorption spectrophotometry (2.4.2), measuring at 767 nm CH3
H3CO O
and using potassium solution AAS, suitably diluted with the CH3
strontium chloride solution, for the standard solutions or,
alternatively by Method A for flame photometry (2.4.4).
1 g of Potassium Chloride is equivalent to 0.5245 g of K. , HCl
For total chloride — Titrate 50 ml of solution A with 0.1 M
silver nitrate using potassium chromate solution as indicator.
O
1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of Cl. N

1 g of Sodium Chloride, and of Potassium Chloride is


equivalent to 0.6066 and 0.4756 g of Cl respectively. C30H35NO3,HCl Mol. Wt. 493.5
For citrate — Weigh accurately about 2.0 g and dissolve in Ormeloxifen Hydrochloride is trans-7-methoxy-2,2-dimethyl-
50 ml of anhydrous glacial acetic acid by heating at about 3-phenyl-4[4-(2-pyrrolidinoethoxy)phenyl]chroman
50º. Cool, allow to stand for 10 minutes. Titrate with 0.1 M hydrochloride.
perchloric acid, using 1-naphtholbenzein solution as
indicator. Carry out a blank titration. Ormeloxifen Hydrochloride contains not less than 98.5 per
cent and not more than 101.5 per cent of C30H35NO3,HCl,
1 ml of 0.1 M perchloric acid is equivalent to 0.006303 g of calculated on the dried basis.
C6H5O7.
Description. A white to off-white powder.
1 g of Sodium Citrate is equivalent to 0.6430 g of C6H5O7.
Identification
For dextrose — Weigh accurately about 7.5 g, dissolve in
40 ml of water, add 0.5 ml of dilute ammonia solution, and A. Determine by infrared absorption spectrophotometry (2.4.6).
dilute to 50 ml with water. Mix well, allow to stand for Compare the spectrum with that obtained with ormeloxifen
30 minutes, filter the solution, if turbid, and measure the optical hydrochloride RS or with the reference spectrum of
rotation in a 2-dm tube (2.4.22). The observed rotation, in ormeloxifen.
degrees, multiplied by 0.9477 and 1.0424 represents the weight
B. When examined in the range 230 nm to 360 nm (2.4.7), a
in g of C6H12O6 and C6H12O6,H2O respectively, as appropriate,
0.01 per cent w/v solution in methanol shows absorption
in the weight taken for the Assay.
maxima at about 278 and 282 nm.
Storage. Store protected from moisture in sachets, preferably C. Determine by thin-layer chromatography (2.4.17), coating
made of aluminum foil, containing sufficient powder for a single the plate with silica gel G.
dose or for a day’s treatment or for use in hospitals, in bulk
containers containing sufficient quantity to produce a volume Mobile phase. A mixture of 90 volumes of chloroform and
of solution appropriate to the daily requirements of the 10 volumes of methanol.
hospital concerned. Test solution. Dissolve 0.25 g of the substance under
Labelling. The label states (1) for sachets, the total weights, examination in 100 ml of chloroform.
in g, of each constituent; (2) for bulk containers, the weights, Reference solution. A 0.25 per cent w/v solution of ormeloxifen
in g, of each constituent in a stated quantity, in g, of the oral hydrochloride RS in chloroform.
powder; (3) the molar concentration in millimoles per litre of
Apply to the plate 2 µl of each solution. After development,
sodium, potassium, chloride and citrate ions, and of dextrose
dry the plate in air, until the odour of the solvents is no longer
as well as the total osmolar concentration in mOsmol per litre
detectable and spray with a 0.3 per cent w/v solution of
of the solution prepared from the oral powder; (4) the total
potassium permanganate. The principal spot in the
weight of the contents of the container; (5) the directions for
chromatogram obtained with the test solution corresponds to
use; (6) that any portion of the solution prepared from the oral
that in the chromatogram obtained with the reference solution.
powder that remains unused for 24 hours after preparation
should be discarded; (7) the storage conditions. g, of the oral D. To 1 ml of a 2 per cent w/v solution in ethanol (95 per cent)
powder. add 1 ml of a saturated solution of picric acid in water, stir

862
IP 2007 ORMELOXIFEN HYDROCHLORIDE TABLETS

well and set aside for 5 minutes. The yellow precipitate obtained six replicate analyses calculate the content of trans-ormeloxifen
after washing with water and drying at 60º for 4 hours melts at hydrochloride and cis-ormeloxifen hydrochloride in the
212º to 218º (2.4.21). substance under examination by comparing with the peak
responses for trans-ormeloxifen hydrochloride and cis-
Tests ormeloxifen hydrochloride respectively obtained with
Total basic substances. Weigh accurately 0.5 g, dissolve in reference solution.
25 ml of anhydrous glacial acetic acid, add 10 ml of a 5 per Storage. Store protected from moisture.
cent w/v solution of mercuric acetate in anhydrous glacial
acetic acid. Titrate with 0.1 M perchloric acid, using crystal
violet solution as indicator to a bluish green end-point. Carry
out a blank titration. Ormeloxifen Hydrochloride Tablets
1 ml of 0.1 M perchloric acid is equivalent to 0.04935 g of Centchroman Hydrochloride Tablets; Centchroman
C30H35NO3,HCl. Tablets
cis-Isomer. Not more than 1.5 per cent of the total content of Ormeloxifen Hydrochloride Tablets contain not less than
hydrochlorides of trans-and cis-isomers determined in the 90.0 per cent and not more than 110.0 per cent of the stated
Assay. amount of Ormeloxifen hydrochloride, C30H35NO3,HCl.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Identification
Loss on drying (2.4.19). Not more than 3.5 per cent, determined Shake a quantity of the powdered tablets containing 0.1 g of
on 1.0 g by drying in an oven at 105º for 4 hours. Ormeloxifen Hydrochloride with 10 ml of chloroform, filter
Assay. Determine by liquid chromatography (2.4.14). and evaporate the filtrate to dryness at a pressure not
exceeding 0.7 kPa. The residue complies with the following
Test solution. A 0.004 per cent w/v solution of the substance
tests.
under examination in the mobile phase.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Reference solution. A solution containing 0.004 per cent w/v
Compare the spectrum with that obtained with ormeloxifen
of trans-ormeloxifen hydrochloride RS and 0.0006 per cent
hydrochloride RS or with the reference spectrum of
w/v of cis-ormeloxifen hydrochloride RS in the mobile phase.
ormeloxifen.
Chromatographic system B. When examined in the range 230 nm to 360 nm (2.4.7), a
– a stainless steel column 25 cm x 4.6 mm, packed with 0.01 per cent w/v solution in methanol shows absorption
octadecylsilane bonded to porous silica (5 µm), maxima at about 278 and 282 nm.
– mobile phase: 80 volumes of acetonitrile and 20 volumes
of water containing 0.04 per cent w/v solution of C. Determine by thin-layer chromatography (2.4.17), coating
tetramethylammonium hydroxide adjusted to pH 7.6 the plate with silica gel G.
with phosphoric acid, Mobile phase. A mixture of 90 volumes of chloroform and
– flow rate. 1.5 ml per minute, 10 volumes of methanol.
– spectrophotometer set at 280 nm, Test solution. Dissolve 0.25 g of the substance under
– a 20 µl loop injector. examination in 100 ml of chloroform.
Inject the reference solution and use an attenuation such that Reference solution. A 0.25 per cent w/v solution of ormeloxifen
the response for the principal peak due to trans-ormeloxifen hydrochloride RS in chloroform.
hydrochloride is more than 50 per cent of the full-scale
deflection of the recorder. Make a total of six injections. When Apply to the plate 2 µl of each solution. After development,
the chromatograms are recorded under the conditions dry the plate in air, until the odour of the solvents is no longer
described above, trans-ormeloxifen hydrochloride is eluted detectable and spray with a 0.3 per cent w/v solution of
before cis-ormeloxifen hydrochloride.The test is not valid potassium permanganate. The principal spot in the
unless the relative standard deviation of six replicate injections chromatogram obtained with the test solution corresponds to
is not greater than 6 per cent and the resolution between the that in the chromatogram obtained with the reference solution.
peaks due to cis-ormeloxifen hydrochloride and trans- Tests
ormeloxifen hydrochloride is greater than 1. If necessary, adjust
the proportions and the flow rate of the mobile phase to obtain Dissolution (2.5.2).
proper resolution. Inject a suitable volume of test solution Apparatus. No 2
and record the response. From the average peak areas of the Medium. 900 ml of water.

863
ORPHENADRINE CITRATE IP 2007

Speed and time. 100 rpm and 45 minutes. hydrochloride and cis-ormeloxifen hydrochloride in the
Withdraw a suitable volume of the medium and filter. Measure substance under examination.
the absorbance of the filtrate, suitably diluted if necessary, at Storage. Store protected from moisture.
the maximum at about 278 nm (2.4.7). Calculate the content of
C30H35NO3,HCl in the medium from the absorbance obtained
from a solution of known concentration of Ormeloxifen
hydrochloride RS in water. Orphenadrine Citrate
D. Not less than 70 per cent of the stated amount of
CH3
C30H35NO3,HCl.
N
cis-Isomer. Not more than 1.5 per cent of the total content of CH3 O CH3
HO COOH
hydrochlorides of trans-and cis-isomers determined in the ,
Assay. HOOC COOH

Other Tests. Comply with the tests stated under Tablets.


Assay. Determine by liquid chromatography (2.4.17).
C18H23NO,C6H807 Mol.461.5
Test solution. Weigh and powder 20 tablets. Shake a quantity
Orphenadrine Citrate is (RS)-dimethyl[2-(2-
of the powder containing 0.1 g of Ormeloxifen Hydrochloride
methylbenzhydryloxy)ethyl]amine dihydrogen citrate.
with three quantities, each of 5 ml, of methanol, centrifuge
each extract, dilute the combined extracts to 25 ml with Orphenadrine Citrate contains not less than 98.5 per cent and
methanol and then dilute 250 µl of the resulting solution to 25 not more than 101.0 per cent of C18H23NO, C6H807, calculated
ml with the mobile phase. on the dried basis.
Reference solution. A solution containing 0.004 per cent w/v Description. A white or almost white, crystalline powder;
of trans-ormeloxifen hydrochloride RS and 0.0006 per cent odourless or almost odourless.
w/v of cis-ormeloxifen hydrochloride RS in the mobile phase.
Identification
Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with Test A may be omitted if tests B, C, D and E are carried out.
octadecylsilane bonded to porous silica (5 µm), Tests B, C and D may be omitted if tests A and E are carried
– mobile phase: 80 volumes of acetonitrile and 20 volumes out.
of water containing 0.04 per cent w/v solution of A. Determine by infrared absorption spectrophotometry,(2.4.6).
tetramethylammonium hydroxide adjusted to pH 7.6 Compare the spectrum with that obtained with orphenadrine
with phosphoric acid, citrate RS.
– flow rate. 1.5 ml per minute,
B. When examined in the range 230 nm to 360 nm (2.4.7), a
– spectrophotometer set at 280 nm,
0.06 per cent w/v solution in ethanol (95 per cent) shows
– a 20 µl loop injector. absorption maxima at 258 nm, 264 nm and 271 nm; absorbances
Inject the reference solution and use an attenuation such that at the maxima are about 0.68, 0.72 and 0.47 respectively.
the response for the principal peak due to trans-ormeloxifen C. Dissolve 50 mg in 10 ml of ethanol (50 per cent), add 10 ml
hydrochloride is more than 50 per cent of full-scale deflection of picric acid solution and allow to stand. The precipitate.
of the recorder. Make a total of six injections. When the after recrystallisation from ethanol (95 per cent), melts at
chromatograms are recorded under the conditions described about 89º or at about 107º (2.4.21).
above, trans-ormeloxifen hydrochloride is eluted before cis-
ormeloxifen hydrochloride.The test is not valid unless the D. Dissolve 5 mg in 2 ml of sulphuric acid; an orange-red
relative standard deviation of six replicate injections is not colour is produced.
greater than 6 per cent and the resolution between the peaks E. To 1 g add 10 ml water and 2 ml of 5 M sodium hydroxide,
due to cis-ormeloxifen hydrochloride and trans-ormeloxifen shake with two quantities, each of 10 mI, of chloroform and
hydrochloride is greater than 1.If necessary, adjust the discard the chloroform. Heat the aqueous solution to boiling
proportions and the flow rate of the mobile phase to obtain with an excess of mercuric sulphate solution, filter if necessary
proper resolution. Inject a suitable volume of test solution and boil the resulting solution with 0.2 ml of dilute potassium
and record the response. From the average peak areas of the permanganate solution; the solution is decolorised and a
six replicate analyses calculate the content of trans-ormeloxifen white precipitate is produced.

864
IP 2007 ORPHENADRINE HYDROCHLORIDE

Tests Orphenadrine Hydrochloride


Quaternary ammonium salt. Determine by thin-layer C18H23NO,HCl Mol. Wt.305.8
chromatography (2.4.17), coating the plate with silica gel G.
Orphenadrine Hydrochloride is (RS)-dimethyl [2-
Mobile phase. A mixture of 50 volumes of 2-propanol, (2methylbenzhydryloxy) ethyl]amine hydrochloride.
30 volumes of butyl acetate. 15 volumes of water and
5 volumes of strong ammonia solution. Orphenadrine Hydrochloride contains not less than 98.5 per
cent and not more than 101.0 per cent of C18H23NO,HCl.
Test solution. Dissolve 0.1 g of the substance under calculated on the dried basis.
examination in 10 ml methanol.
Description. A white or almost white, crystalline powder;
Reference solution. A 0.005 per cent w/v solution of odourless or almost odourless.
ethyldimethyl [2-(2-methylbenzhydryloxy)ethyl ]ammonium
chloride RS. Identification
Apply to the plate 5 µl of each solution. After development, Tests A and F may be omitted if tests B, C, D and E are carried
dry the plate in air and spray with dilute potossium out. Tests B, C and D may be omitted if tests A, E and F are
iodobismuthate solution. Any spot corresponding to carried out.
ethyldimethyl [2-(2-methylbenzhydryloxy) ethyl] ammonium
chloride in the chromatogram obtained with the test solution A. Determine by infrared absorption spectrophotometry,(2.4.6).
is not more intense than the spot in the chromatogram obtained Compare the spectrum with that obtained with orphenadrine
with the reference solution. hydrochloride RS.
Secondary amine. Determine by thin-layer chromatography B. When examined in the range 230 nm to 360 nm (2.4.7), a
(2.4.17) coating the plate with silica gel GF254. 0.06 per cent w/v solution in ethanol (95 per cent) shows
three absorption maxima. at about 258 nm,264 nm and 271 nm;
Mobile phase. A mixture of 96 volumes of 1-butanol and
absorbances at the maxima, about 1.07, 1.13 and 0.73
4 volumes of strong ammonia solution.
respectively.
Test solution. Dissolve 0.4 g of the substance under
examination in 10 ml methanol. C. Dissolve about 5 mg in 2 ml of sulphuric acid; an orange-
red colour is produced.
Reference solution. A 0.02 per cent w/v solution of methyl [2-
(2-methylbenzhydryloxy)ethyl]amine hydrochloride RS. D. Dissolve about 50 mg in 10 ml of ethanol (95per cent), add
10 ml of picric acid solution and allow to stand. The
Apply to the plate 10 µl of each solution. After development, precipitate, after recrystallisation from ethanol (95 per cent)
dry in air and examine in ultra-violet light at 254 nm. Spray the melts at about 89º or at about 107º (2.4.21).
plate with dilute potassium iodobismuthate solution and
examine again. By each method of visualisation any spot Tests
corresponding to methyl [2-(2-methylbenzhydryloxy)
ethyl]amine hydrochloride in the chromatogram obtained with Quaternary ammonium salt. Determine by thin-layer
the test solution is not more intense than the spot in the chromatography (2.4.17), coating the plate with silica gel G.
chromatogram obtained with the reference solution. Mobile phase. A mixture of 50 volumes of 2-propanol,
Heavy Metals (2.3.13). 2.0 g complies with the limit test for 30 volumes of butyl acetate. 15 volumes of water and
heavy metals, Method B (10 ppm). Use 2 ml of lead standard 5 volumes of strong ammonia solution.
solution (10 ppm Pb) to prepare the standard. Test solution. Dissolve 0.1 g of the substance under
Sulphated ash (2.3.18). Not more than 0.1 per cent. examination in 10 of methanol.
Loss on drying (2.3.19). Not more than 0.5 per cent, determined Reference solution. A 0.005 per cent w/v solution of
on 1.0 g by drying in an oven at 105° for 3 hours. ethyldimethyl [2-(2-methylbenzhydryloxy) ethyl] ammonium
chloride RS.
Assay. Weigh accurately about I.0 g dissolve in 30 ml of
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric Apply to the plate 5 µl of each solution. After development,
acid, determining the end-point potentiometrically (2.4.25). dry the plate in air and spray with dilute potossium
Carry out a blank titration. iodobismuthate solution. Any spot corresponding to
ethyldimethyl [2-(2-methylbenzhydryloxy) ethyl] ammonium
1 ml of 0.1 M perchloric acid is equivalent to 0.04615 g of
chloride in the chromatogram obtained with the test solution
C18H23NO, C6H807.
is not more intense than the spot in the chromatogram obtained
Storage. Store protected from light. with the reference solution.

865
ORPHENADRINE TABLETS IP 2007

Secondary amine. Determine by thin-layer chromatography B. Dissolve 50 mg in 10 ml of ethanol (50 per cent), add 10 ml
(2.4.17) coating the plate with silica gel GF254. of picric acid solution and allow to stand. The precipitate.
after recrystallisation from ethanol (95 per cent), melts at
Mobile phase. A mixture of 96 volumes of l-butanol and
about 89º or at about 107º (2.4.21).
4 volumes of strong ammonia solution.
C. A 5 per cent w/v solution gives reaction A of chlorides
Test solution. Dissolve 0.4 g of the substance under
(2.3.1).
examination in 10 methanol.
Reference solution. A 0.02 per cent w/v solution of methyl [2- Tests
(2-methylbenzhydryloxy)ethyl]amine hydrochloride RS.
Other tests. Comply with the tests stated under Tablets.
Apply to the plate 10 µl of each solution. After development,
Assay. Weigh and powder 20 tablets. Weigh accurately a
dry in air and examine in ultra-violet light at 254 nm. Spray the
quantity of the powder containing about 70 mg Orphenadrine
plate with dilute potassium iodobismuthate solution and
Hydrochloride and dissolve as completely as possible in a
examine again. By each method of visualisation any spot
mixture of 5 ml of water and 5 ml of 2 M hydrochloric acid.
corresponding to methyl [2-(2-methylbenzhydryloxy)
Without delay extract with four quantities, each of 15 ml, of
ethyl]amine hydrochloride in the chromatogram obtained with
chloroform, filter the combined extracts and evaporate to about
the test solution is not more intense than the spot in the
20 ml. Add 30 ml of anhydrous glacial acetic and 2 ml of
chromatogram obtained with the reference solution.
mercuric acetate solution and titrate with 0.02 M perchloric
Heavy Metals (2.3.13). 2.0 g complies with the limit test for acid determining the end point potentiometrically (2.4.25).
heavy metals, Method B (10 ppm). Use 2 ml of lead standard Carry out a blank titration.
solution (10 ppm Pb) to prepare the standard. 1 ml of O.02 M perchloric acid is equivalent to 0.006117 g of
Sulphated ash (2.3.18). Not more than 0.1 per cent. C18H23NO,HCl.
Loss on drying (2.3.19). Not more than 0.5 per cent, determined Storage. Store protected from light and moisture.
on 1.0 g by drying in an oven at 105° for 3 hours.
Assay. Weigh accurately about 1.0 g, dissolve in 20 ml of
anhydrous glacial acetic acid, add 20 ml of mercuric acetate
solution and titrate with 0.1 M perchloric acid using Oseltamivir Phosphate
1-naphtholbenzein solution as indicator. Carry out a blank
titration. H3C
1 ml of 0.1 M perchloric acid is equivalent to 0.03058 g of H3C
C18H23NO, HCl.
O
H O CH3 , H3PO4
Storage. Store protected from light.
N
H3C H2N O
O

Orphenadrine Tablets C16H28N2O4, H3PO4 Mol. Wt. 410.4


Orphenadrine Hydrochloride Tablets Oseltamivir Phosphate is phosphoric acid salt of ethyl
(3R,4R,5S)-4-(acetylamino)-5-amino-3-(1-ethylpropoxy)-1-
Orphenadrine Tablets contain not less than 92.5 per cent and
cyclohexene-1-carboxylate.
not more than 107.5 per cent of the stated amount of
orphenadrine. hydrochloride, C18H23NO,HCl. Oseltamivir Phosphate contain not less than 98.0 per cent and
not more than 102.0 per cent of C16H28N2O4 .H3PO4, calculated
Identification on the dried basis.
Extract a quantity of the powdered tablets containing about Description. A white to off-white powder.
0.15 g of Orphenadrine Hydrochloride with chloroform, filter
and evaporate the filtrate to dryness. The residue complies Identification
with the following tests.
A. Determine by infrared absorption spectrophotometry (2.4.6).
A. Dissolve 5 mg in 2 ml of sulphuric acid; an orange-red Compare the spectrum with that obtained with oseltamivir
colour is produced. phosphate RS.

866
IP 2007 OSELTAMIVIR CAPSULES

B. In the Assay, the principal peak in the chromatogram Loss on drying (2.4.19). Not more than 0.5 per cent, determined
obtained with the test solution corresponds to the peak in the on 1 g by drying in an oven at 105º.
chromatogram obtained with the reference solution. Assay. Determine by liquid chromatography (2.4.14).
Tests Test solution. Dissolve 50 mg of the substance under
examination in 50.0 ml of the mobile phase. Dilute 10.0 ml of
Specific optical rotation (2.4.22). - 42.0º to - 48.0º, determined
the solution to 50.0 ml with the mobile phase.
in a 1.0 per cent w/v solution in methanol.
Related substances. Determine by liquid chromatography Reference solution. A 0.02 per cent w/v solution of oseltamivir
(2.4.14). phosphate RS in the mobile phase.

NOTE — Prepare the solutions immediately before use. Chromatographic system


– a stainless steel column 15 cm x 4.6 mm, packed with
Test solution. Dissolve 25 mg of the substance under octylsilane bonded to porous silica (5 µm) (such as YMC
examination in 25 ml of the mobile phase. pack PRO C8),
Reference solution (a). A 0.1 per cent w/v solution of – column temperature 50º,
oseltamivir phosphate RS in the mobile phase. – mobile phase: a mixture of 66 volumes of buffer solution
prepared by dissolving 6.8 g of anhydrous monobasic
Reference solution (b). Dilute 1 ml of reference solution (a) to
potassium phosphate in 1000 ml of water, adjusting the
100 ml with the mobile phase.
pH to 6.0 with dilute sodium hydroxide, 24.5 volumes
Chromatographic system of methanol and 23.5 volumes of acetonitrile,
– a stainless steel column 15 cm x 4.6 mm, packed with – flow rate. 1 ml per minute,
octylsilane bonded to porous silica (5 µm) (such as YMC – spectrophotometer set at 207 nm,
pack PRO C8), – a 20 µl loop injector.
– column temperature 50º,
Inject the reference solution. The test is not valid unless the
– mobile phase: a mixture of 66 volumes of a buffer solution
tailing factor is not more than 2.0, the column efficiency in not
prepared by dissolving 6.8 g of anhydrous monobasic
less than 2000 theoretical plates and the relative standard
potassium phosphate in 1000 ml of water and adjusting
deviation for replicate injections is not more than 2.0 per cent.
the pH to 6.0 with dilute sodium hydroxide, 24.5 volumes
of methanol and 23.5 volumes of acetonitrile, Inject the test solution and the reference solution.
– flow rate. 1 ml per minute, Calculate the content of C16H28N2O4,H3PO4.
– spectrophotometer set at 207 nm,
– a 20 µl loop injector. Storage. Store protected from light and moisture.

Inject reference solution (a). The test is not valid unless the
column efficiency is not less than 2000 theoretical plates and
the tailing factor is not more than 2.0.
Oseltamivir Capsules
Inject the test solution and reference solution (b). In the Oseltamivir Phosphate Capsules
chromatogram obtained with the test solution, the area of any Oseltamivir Capsules contain Oseltamivir Phosphate.
secondary peak is not more than 0.5 time the area of the peak
Oseltamivir Capsules contain not less than 90.0 per cent and
in the chromatogram obtained with the reference solution (b)
not more than 110.0 per cent of the stated amount of
(0.5 per cent) and the sum of all the secondary peaks is not
oseltamivir, C16H28N2O4.
more than the area of the peak in the chromatogram obtained
with the reference solution (b) (1.0 per cent). Identification
Phosphoric acid. 23.4 to 24.4 per cent, calculated on a dried
In the Assay, the principal peak in the chromatogram obtained
basis.
with the test solution corresponds to the peak in the
Weigh accurately about 0.2 g and dissolve in 40 ml of distilled chromatogram obtained with the reference solution.
water. Titrate with 0.1 M sodium hydroxide, determining the
end-point potentiometrically (2.4.25). Carry out a blank titration. Tests
1 ml of 0.1 M sodium hydroxide is equivalent to 0.0049 g of Dissolution (2.5.2).
phosphoric acid. Apparatus. No. 1
Heavy metals (2.3.13). 1 g complies with the limit test for heavy Medium. 900 ml of 0.1 M hydrochloric acid.
metals, Method A (20 ppm). Speed and time. 50 rpm and 45 minutes

867
OSELTAMIVIR ORAL SUSPENSION IP 2007

Withdraw a suitable volume of the medium and filter. Determine Test solution. Weigh accurately a quantity of the mixed
by liquid chromatography (2.4.14). contents of 20 capsules containing about 200 mg of oseltamivir,
Test solution. Use the filtrate. disperse in 100.0 ml of the mobile phase and filter. Dilute 5.0 ml
of the solution to 50.0 ml with the mobile phase.
Reference solution. Dissolve 50 mg of oseltamivir phosphate
Reference solution. A 0.02 per cent w/v solution of oseltamivir
RS in 20 ml of the mobile phase and dilute to 100 ml with the
phosphate RS in the mobile phase.
dissolution medium. Dilute 5 ml of the solution to 25 ml with
the dissolution medium. Chromatographic system
– a stainless steel column 15 cm x 4.6 mm, packed with
Use the chromatographic system described under Assay.
octylsilane bonded to porous silica (5 µm),
D. Not less than 80 per cent of the stated amount of – mobile phase: a mixture of 66 volumes of a buffer solution
C16H28N2O4. prepared by dissolving 6.8 g of anhydrous monobasic
Related substances. Determine by liquid chromatography potassium phosphate in 1000 ml of water and adjusting
(2.4.14). the pH to 6.0 with dilute sodium hydroxide, 24.5 volumes
of methanol and 23.5 volumes of acetonitrile,
NOTE — Prepare the solutions immediately before use. – flow rate. 1.2 ml per minute,
Test solution. Weigh accurately a quantity of the mixed – spectrophotometer set at 207 nm,
contents of 20 capsules containing about 50 mg of oseltamivir, – a 20 µl loop injector.
disperse in 50 ml of mobile phase and filter. Inject the reference solution. The test is not valid unless the
tailing factor is not more than 2.0, the column efficiency in not
Reference solution (a). A 0.1 per cent w/v solution of less than 2000 theoretical plates and the relative standard
oseltamivir phosphate RS in the mobile phase. deviation for replicate injections is not more than 2.0 per cent.
Reference solution (b). Dilute 1 ml of reference solution to Inject the test solution and the reference solution.
100 ml with the mobile phase.
Calculate the content of C16H28N2O4.
Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with Storage. Store protected from moisture, at a temperature not
octylsilane bonded to porous silica (5 µm) (such as YMC exceeding 30º.
pack PRO C8), Labelling. The label states the strength in terms of the
– mobile phase: a mixture of 66 volumes of a buffer solution equivalent amount of oseltamivir.
prepared by dissolving 6.8 g of anhydrous monobasic
potassium phosphate in 1000 ml of water, adjusting the
pH to 6.0 with dilute sodium hydroxide, 24.5 volumes
of methanol and 23.5 volumes of acetonitrile,
Oseltamivir Oral Suspension
– flow rate. 1.5 ml per minute, Oseltamivir Phosphate Oral Suspension
– spectrophotometer set at 207 nm,
Oseltamivir Oral Suspension is a mixture consisting of
– a 20 µl loop injector.
Oseltamivir Phosphate with buffering agents and other
Inject reference solution (a). The test is not valid unless the excipients. It contains a suitable flavouring agent. It is filled in
column efficiency is not less than 2000 theoretical plates and a sealed container.
the tailing factor is not more than 2.0.
The suspension is constituted by dispersing the contents of
Inject the test solution and reference solution (b). In the the sealed container in the specified volume of water just
chromatogram obtained with the test solution, the area of any before issue.
secondary peak is not more than the area of the peak in the
Oseltamivir Oral Suspension contains not less than 90.0 per
chromatogram obtained with the reference solution (b) (1.0
cent and not more than 110.0 per cent of the stated amount of
per cent) and the sum of the areas of all the secondary peaks
oseltamivir C16H28N2O4.
is not more than twice the area of the peak in the chromatogram
obtained with the reference solution (b) (2.0 per cent). The contents of the sealed container comply with the
following requirement.
Other tests. Comply with the tests stated under Capsules.
Water (2.3.43). Not more than 2.5 per cent, determined on
Water (2.3.43). Not more than 5.0 per cent, determined on 0.5 g.
1.0 g.
Assay. Determine by liquid chromatography (2.4.14).
The constituted suspension complies with the requirements
NOTE — Prepare the solutions immediately before use. stated under Oral liquids and with the following requirements.

868
IP 2007 OXAZEPAM

Identification Test solution. Weigh accurately a quantity of the suspension


containing 60 mg of oseltamivir, dissolve in 250.0 ml of the
A. In the Assay, the principal peak in the chromatogram mobile phase and filter. Dilute 10.0 ml of the solution to 25.0 ml
obtained with the test solution corresponds to the peak in the with the mobile phase and filter.
chromatogram obtained with reference solution (b).
Reference solution. A 0.0125 per cent w/v solution of
B. To 2 ml of the reconstituted suspension, add 2 ml of dilute oseltamivir phosphate RS in the mobile phase.
nitric acid and 4 ml of a 10 per cent w/v solution of ammonium
molybdate and warm the solution. A bright yellow precipitate Chromatographic system
is formed. – a stainless steel column 15 cm x 4.6 mm, packed with
octylsilane bonded to porous silica (5 µm),
Tests – mobile phase: a mixture of 60 volumes of a buffer solution
prepared by dissolving 6.8 g of potassium dihydrogen
pH (2.4.24). 3.0 to 5.0.
phosphate in 1000 ml of water and adjusting the pH to
Related substances. Determine by liquid chromatography 6.0 with dilute sodium hydroxide, 24.5 volumes of
(2.4.14). methanol and 23.5 volumes of acetonitrile,
NOTE — Prepare the solutions immediately before use. – flow rate. 1.2 ml per minute,
– spectrophotometer set at 207 nm,
Test solution. Weigh accurately a quantity of the suspension – a 20 µl loop injector.
containing 25 mg of oseltamivir, dissolve in 25 ml of the mobile
phase and filter. Inject the reference solution. The test is not valid unless the
tailing factor is not more than 2.0, the column efficiency in not
Reference solution (a). A 0.1 per cent w/v solution of less than 2000 theoretical plates and the relative standard
oseltamivir phosphate RS in the mobile phase. deviation for replicate injections is not more than 2.0 per cent.
Reference solution (b). Dilute 1 ml of reference solution (a) to Inject the test solution and the reference solution.
100 ml with the mobile phase.
Calculate the content of C16H28N2O4.
Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with Storage. Store protected from moisture, at a temperature not
octylsilane bonded to porous silica (5 µm), exceeding 30°.
– column temperature 35º, Labelling. The label states (1) the quantity of active ingredient
– mobile phase: 70 volumes of a buffer solution prepared in terms of the equivalent amount of oseltamivir; (b) the
by dissolving 6.8 g of potassium dihydrogen phosphate temperature of storage and the period during which the
in 1000 ml of water and adjusting the pH to 6.0 with constituted suspension may be expected to be satisfactory
dilute sodium hydroxide, 15 volumes of methanol and for use.
15 volumes of acetonitrile,
– flow rate. 1.5 ml per minute,
– spectrophotometer set at 207 nm,
– a 20 µl loop injector. Oxazepam
Inject reference solution (a). The test is not valid unless the
H O
column efficiency is not less than 2000 theoretical plates and N
the tailing factor is not more than 2.0.
OH
Inject the test solution and reference solution (b). In the N
Cl
chromatogram obtained with the test solution, the area of any
secondary peak is not more than the area of the peak in the
chromatogram obtained with the reference solution (b)
(1.0 per cent) and the sum of areas of all the secondary peaks
is not more than 3 times the area of the peak in the
C15HIICIN2O2 Mol. Wt. 286.7
chromatogram obtained with the reference solution (b)
(3.0 per cent). Oxazepam is 7-chloro-3-hydroxy-5-phenyl-1,3-dihydro-2H-
1,4-benzodiazepin-2-one.
Other tests. Complies with the tests stated under Oral liquids.
Oxazepam contains not less than 98.5 per cent and not more
Assay. Determine by liquid chromatography (2.4.14).
than 101.0 per cent of C15HIICIN2O2, calculated on the dried
NOTE — Prepare the solutions immediately before use. basis.

869
OXAZEPAM TABLETS IP 2007

Description. A white or almost white, crystalline powder. Reference solution (c). Dilute 1 ml of reference solution (a) to
100 ml with acetone.
Identification Reference solution (d). Dilute 5 ml of solution (d) to 10 ml with
Test A may be omitted if tests B, C and D are carried out. Tests acetone.
B and D may be omitted if tests A and C are carried out. Apply to the plate 20 µl of each solution. Allow the mobile
A. Determine by infrared absorption spectrophotometry (2.4.6). phase to rise 17 cm in the same direction as the washing with
Compare the spectrum with that obtained with oxazepam RS. methanol. Dry in air and examine in ultraviolet light at 254 nm.
Any secondary spot in the chromatogram obtained with test
B. Prepare the solutions immediately before use, protected solution (a) is not more intense than the spot in the-
from light. chromatogram obtained with reference solution (c) and at most
Dissolve about 20 mg in sufficient ethanol (95 per cent) to one such spot is more intense than the spot in the
produce 100 ml. Dilute 10 ml of the solution to 50 ml with chromatogram obtained with reference solution (d). The test
ethanol (95 per cent) (solution A). Dilute 10 ml of solution A is not valid unless the chromatogram obtained with reference
to 100 ml with ethanol (95 per cent) (solution B). solution (b) shows two clearly separated spots.

When examined in the range 230 nm to 360 nm (2.4.7), solution Sulphated ash (2.3.18). Not more than 0.1 per cent.
A shows an absorption maximum at about 316 nm. When Loss on drying (2.3.19). Not more than 0.5 per cent, determined
examined in the range 220 nm to 250 nm, solution B shows an on 1.0 g by drying in an oven at 105º at a pressure not exceeding
absorption maximum at about 229 nm; absorbance at about 0.7 kPa.
229 nm, 1.220 to 1.300.
Assay. Weigh accurately about 0.25 g and dissolve in a mixture
C. In the test for Related substances, the principal spot in the of 10 ml of anhydrous glacial acetic acid and 90 ml of acetic
chromatogram obtained with test solution (b) corresponds to anhydride and titrate with 0.1 M perchloric acid determining
that in the chromatogram obtained with reference solution the end point potentiometrically (2.4.25). Carry out a blank
(b). titration.
D. Dissolve about 20 mg in a mixture of 5 ml of hydrochloric 1 ml of 0.1 M perchloric acid is equivalent to 0.02867 g of
acid and 10 ml of water. Heat to boiling for 5 minutes and cool. Cl5HIICIN2O2.
Add 2 ml of a 0.1 per cent w/v solution of sodium nitrite and Storage. Store protected from light and moisture.
allow to stand for 1 minute. Add 1 ml of a 0.5 per cent w/v
solution of sulphamic acid, mix and allow to stand for 1 minute.
Add 1 ml of 0.1 per cent w/v solution of a N- (1-naphthyl)
ethylenediamine dihydrochloride; a red colour is produced. Oxazepam Tablets
Oxazepam Tablets contain not less than 90.0 per cent and not
Tests more than 110.0 per cent of the stated amount of oxazepam,
CI5H11CIN2O2.
Related substances: Carry out the test protected from light.
Determine by thin-layer chromatography (2.4.17), coating the Identification
plate with silica gel GF254.
A. Extract a quantity of the powdered tablets containing
Wash the plate with methanol before use. 20 mg of Oxazepam with 25 ml of chloroform, filter, evaporate
Mobile phase. A mixture of 100 volumes of dichloromethane to dryness and dry the residue at 60° at a pressure not exceeding
and 10 volumes of methanol. 0.7 kPa.

Test solution (a). Dissolve 50 mg of the substance under On the residue, determine by infrared absorption
examination in sufficient acetone to produce 10 ml. spectrophotometry (2.4.6). Compare the spectrum with that
obtained with oxazepam RS or with the reference spectrum of
Test solution (b). Dilute 2 ml of test solution (a) to 10 ml with oxazepam.
acetone.
B. When examined in the range 210 nm to 360 nm (2.4.7), the
Reference solution (a). Dissolve 10 mg of oxazepam RS in solution obtained in the Assay shows absorption maxima at
sufficient acetone to produce 10 ml. about 230 nm and 316 nm.
Reference solution (b). Dissolve 10 mg each of oxazepam RS
Tests
and bromazepam RS in sufficient acetone to produce
10 ml. Related substances. Carry out the test protected from light.

870
IP 2007 OXPRENOLOL HYDROCHLORIDE

Determine by thin-layer chromatography (2.4.17) coating the Oxprenolol Hydrochloride contains not less than 98.5 per cent
plate with silica gel GF 254. and not more than 101.5 per cent of C15H23NO3,HCl, calculated
Wash the plate with methanol before use. on the dried basis.

Mobile phase. A mixture of 100 volumes of dichloromethane Description. A white or almost white, crystalline powder.
and 10 volumes of methanol. Identification
Test solution. Shake a quantity of powdered tablets containing
30 mg of Oxazepam with 6 ml of acetone and centrifuge. Test A may be omitted if tests B and C are carried out. Test B
may be omitted if tests A and C are carried out.
Reference solution (a). Dilute 1 volume of the test solution to
100 volumes with acetone. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with oxprenolol RS
Reference solution (b). Dilute 1 volume of the test solution to or with the reference spectrum of oxprenolol.
500 volumes with acetone.
B. In the test for Related substances, the principal spot in the
Reference solution (c). A solution containing 0.1 per cent chromatogram obtained with test solution (b) corresponds to
w/v each of oxazepam RS and bromazepam RS. that in the chromatogram obtained with reference solution (c).
Apply to the plate 20 µl of each of the solutions. Allow the C. Gives reaction A of chlorides (2.3.1).
mobile phase to rise 17 cm in the same direction as in the
washing with methanol. Dry in air and examine in ultraviolet Tests
light at 254 nm. Any secondary spot in the chromatogram Appearance of solution. A 10.0 per cent w/v solution in carbon
obtained with the test solution is not more intense than the dioxide-free water is clear (2.4.1), and not more intensely
spot in the chromatogram obtained with reference solution (a) coloured than reference solution GYS6 (2.4.1).
(1 per cent) and not more than one such spot is more intense
than the spot in the chromatogram obtained with reference pH (2.4.24). 4.5 to 6.0, determined in a freshly prepared
solution (b) (0.2 per cent). The test is not valid unless the 10.0 per cent w/v solution.
chromatogram obtained with reference solution (c) shows two Related substances. Determine by thin-layer chromatography
clearly separated spots. (2.4.17), coating the plate with silica gel G.
Other tests. Comply with the tests stated under Tablets. Mobile phase. A mixture of 88 volumes of chloroform,
Assay. Weigh and powder 20 tablets. Weigh accurately a 12 volumes of methanol and 2 volumes of strong ammonia
quantity of the powder containing about 25 mg of Oxazepam solution.
and shake with 150 mI of ethanol (95 per cent) for 30 minutes. Test solution (a). Dissolve 0.5 g of the substance under
Add sufficient ethanol (95 per cent) to produce 250.0 ml and examination in 10 ml of a mixture of 90 volumes of chloroform
centrifuge. Dilute 5.0 ml of the supernatant liquid to 100.0 ml and 10 volumes of methanol.
with the same solvent and measure the absorbance of the Test solution (b). Dissolve 0.5 g of the substance under
resulting solution at the maximum at about 230 nm (2.4.7). examination in 100 ml of a mixture of 90 volumes of chloroform
Calculate the content of CI5HI1CIN2O2 taking 1250 as the and 10 volumes of methanol.
specific absorbance at 230 nm.
Reference solution (a). A 0.02 per cent w/v solution of the
Storage. Store protected from light at a temperature not substance under examination in a mixture of 90 volumes of
exceeding 30º. chloroform and 10 volumes of methanol.
Reference solution (b). A 0.01 per cent w/v solution of the
substance under examination in a mixture of 90 volumes of
Oxprenolol Hydrochloride chloroform and 10 volumes of methanol.
Reference solution (c). A 0.5 per cent w/v solution of
OH H oxprenolol hydrochloride RS in a mixture of 90 volumes of
O N CH3 chloroform and 10 volumes of methanol.
, HCl Apply to the plate 2 µl of each solution. Allow the mobile
CH2 CH3
O phase to rise 13 cm. Dry the plate in warm air for 10 minutes,
allow to cool, spray with anisaldehyde solution, heat at 105º
C15H23NO3,HCl Mol. Wt. 301.8
for 10 minutes and examine in daylight. Any secondary spot
Oxprenolol Hydrochloride is (RS)-1-(2-allyloxyphenoxy)-3- in the chromatogram obtained with test solution (a) is not
isopropylaminopropan-2-ol hydrochloride. more intense than the spot in the chromatogram obtained

871
OXPRENOLOL TABLETS IP 2007

with reference solution (a) and not more than one such spot is Mobile phase. A mixture of 88 volumes of chloroform,
more intense than the spot in the chromatogram obtained 12 volumes of methanol and 2 volumes of strong ammonia
with reference solution (b). solution.
Heavy metals (2.3.13). 1.0 g complies with the limit test for Test solution. Extract a quantity of the powdered tablets
heavy metals, Method B (20 ppm). containing 0.25 g of Oxprenolol Hydrochloride with 5 ml of
Sulphated ash (2.3.18). Not more than 0.1 per cent. water, centrifuge and use the supernatant liquid.

Loss on drying (2.4.19). Not more than 0.5 per cent, determined Reference solution. Dilute 1 volume of the test solution to
on 1.0 g by drying in an oven at 60º over phosphorus pentoxide 200 volumes with water.
at a pressure of 1.5 to 2.5 kPa for 6 hours. Apply to the plate 2 µl of each solution. Allow the mobile
Assay. Weigh accurately about 0.25 g, dissolve in 60 ml of phase to rise 13 cm. Dry the plate in warm air for 10 minutes,
anhydrous glacial acetic acid, add 5 ml of mercuric acetate allow to cool, spray with anisaldehyde solution, heat at 105º
solution. Titrate with 0.1 M perchloric acid, determining the for 10 minutes and examine in daylight. Any secondary spot
end-point potentiometrically (2.4.25).Carry out a blank in the chromatogram obtained with the test solution is not
titration. more intense than the spot in the chromatogram obtained
with the reference solution.
1 ml of 0.1 M perchloric acid is equivalent to 0.03018 g of
C15H23NO3,HCl. Other Tests. Comply with the tests stated under Tablets.
Storage. Store protected from light and moisture. Assay. Weigh and powder 20 tablets. Weigh accurately a
quantity of the powder containing about 20 mg of Oxprenolol
Hydrochloride, add 25 ml of 0.1 M hydrochloric acid and
sufficient water to produce 250.0 ml. Mix with the aid of
Oxprenolol Tablets ultrasound for 5 minutes, shake for 15 minutes and filter.
Oxprenolol Hydrochloride Tablets Measure the absorbance of the resulting solution at the
maximum at about 273 nm (2.4.7). Calculate the content of
Oxprenolol Hydrochloride Tablets contain not less than C15H23NO3,HCl taking 74.5 as the specific absorbance at
95.0 per cent and not more than 105.0 per cent of the stated 273 nm.
amount of oxprenolol hydrochloride, C15H23NO3,HCl. The
tablets are coated. Storage. Store protected from moisture.

Identification
A.To a quantity of the powdered tablets containing 50 mg of
Oxprenolol Hydrochloride add 10 ml of water and 2 ml of
Oxygen
dilute sodium hydroxide solution, extract with 10 ml of O2 Mol. Wt. 32.0
chloroform and reserve the aqueous layer for test C. Dry the Oxygen contains not less than 99.0 per cent v/v of O2.
chloroform extract over anhydrous sodium sulphate, filter and
evaporate the filtrate to dryness. This monograph applies to oxygen for medicinal use only.
On the residue, determine by infrared absorption Description. A colourless gas; odourless.
spectrophotometry (2.4.6). Compare the spectrum with that
obtained with oxprenolol hydrochloride RS or with the Identification
reference spectrum of oxprenolol hydrochloride. A. A glowing splinter of wood bursts into flame on contact
B. When examined in the range 230 nm to 360 nm (2.4.7), the with the gas.
solution obtained in the Assay shows an absorption maximum B. Shake with alkaline pyrogallol solution; the gas under
only at about 273 nm. examination is absorbed and the solution becoming dark
C. The aqueous layer obtained in test A gives reaction A of brown.
chlorides (2.3.1). C. When tested as described under Assay, not more than
D. The residue obtained in test A melts at about 76º (2.4.21). 1.0 ml of gas remains.

Tests Tests
Related substances. Determine by thin-layer chromatography Carbon dioxide. Not more than 300 ppm v/v, determined by
(2.4.17), coating the plate with silica gel G. using a carbon dioxide detector tube (2.1.1).

872
IP 2007 OXYPHENBUTAZONE

Carbon monoxide. Not more than 5 ppm v/v, determined by compounds and must not be treated with any compound that
using a carbon monoxide detector tube (2.1.1). will be irritating to the respiratory tract when the Oxygen
93 Per Cent is used.
Water vapour. Not more than 67 ppm v/v, determined by using
a water vapour detector tube (2.1.1). Labelling. Label each outlet “Oxygen 93 Per Cent”, when it is
piped directly from the collecting tank to the point of use. If it
Assay (2.3.33). Use 100 ml of the gas under examination and
is stored in cylinders, reduce the pressure by means of a
place spirals of freshly cleaned copper wire and 125 ml of
regulator. Measure the gases with a gas volume meter
ammonia buffer pH 10.9 in the pipette. The volume of the
downstream from the detector tube in order to minimise
residual gas in the burette is not more than 1.0 ml.
contamination or change of the specimens. The shoulder of
Storage. Store under pressure in metal cylinders of the type the cylinder should be painted white and the remainder should
conforming to the appropriate safety regulations. Valves and be painted black. The cylinder should carry a label stating
taps should not be lubricated with oil or grease. “Oxygen 93 Per Cent” and “For medicinal use”.
Labelling. The shoulder of the metal cylinder should be
painted white and the remainder should be painted black. The
cylinder should carry a label stating “Oxygen”. In addition,
“Oxygen” or the symbol “O2” should be stencilled in paint on
Oxyphenbutazone
the shoulder of the cylinder.
OH

Oxygen 93 Per Cent


Oxygen 93 Per Cent contains not less than 90.0 per cent and N , H2O
not more than 96.0 per cent, v/v of O2, the remainder consisting O N
mostly of argon and nitrogen. It is produced from air by the
H3 C
molecular sieve process. O
Description. A colourless gas; odourless.
C19H20N2O3,H2O Mol. Wt. 342.4
Identification
Oxyphenbutazone is (RS)-4-butyl-1-(4-hydroxyphenyl)-2-
A. A glowing splinter of wood bursts into flame on contact phenylpyrazolidine-3,5-dione monohydrate.
with the gas. Oxyphenbutazone contains not less than 98.0 per cent and
B. Shake with alkaline pyrogallol solution; the gas under not more than 101.0 per cent of C19H20N2O3, calculated on the
examination is absorbed and the solution becomes dark brown. anhydrous basis.
C. When tested as described under Assay, not more than Description. A white to yellowish white, crystalline powder;
10.0 ml and not less than 4.0 ml of gas remain. almost odourless.

Tests Identification
Carbon dioxide. Not more than 300 ppm v/v, determined by Test A may be omitted if tests B, C and D are carried out. Tests
using a carbon dioxide detector tube (2.1.1). B and D may be omitted if tests A and C are carried out.
Carbon monoxide. Not more than 5 ppm v/v, determined by A. Determine by infrared absorption spectrophotometry (2.4.6).
using a carbon monoxide detector tube (2.1.1). Compare the spectrum with that obtained with
oxyphenbutazone RS or with the reference spectrum of
Assay (2.3.33). Use 100 ml of the gas under examination and
oxyphenbutazone.
place spirals of freshly cleaned copper wire and 125 ml of
ammonia buffer pH 10.9 in the pipette. The volume of the B. When examined in the range 230 nm to 360 nm (2.4.7), a
residual gas in the burette is not more than 10.0 ml and not 0.001 per cent w/v solution in 0.01 M sodium hydroxide shows
less than 4.0 ml. an absorption maximum at about 254 nm; absorbance at about
254 nm, 0.71 to 0.77.
Storage. Store in cylinders or in a low pressure collecting
tank. Containers used for Oxygen 93 Per Cent must not be C. To 2 ml of a 1 per cent w/v solution in ethanol (95 per cent)
treated with any toxic, sleep-inducing or narcosis-producing add 2 ml of a 0.1 per cent w/v solution of 2,6-dichloroquinone-

873
OXYPHENBUTAZONE TABLETS IP 2007

4-chlorimide in ethanol (95 per cent) and 1 ml of sodium Oxyphenbutazone Tablets


carbonate solution; an intense green colour is produced.
Oxyphenbutazone Tablets contain not less than 92.5 per cent
D. To 0.1 g add 1 ml of glacial acetic acid and 2 ml of and not more than 107.5 per cent of the stated amount of
hydrochloric acid and boil under a reflux condenser for oxyphenbutazone, C19H20N2O3,H2O. The tablets are coated.
30 minutes. Cool, add 10 ml of water and filter. To the filtrate
add 3 ml of a 0.7 per cent w/v solution of sodium nitrite; a Identification
yellow colour develops. To 1 ml of the resulting solution add
a solution of 10 mg of 2-naphthol in 5 ml of sodium carbonate Extract a quantity of the powdered tablets containing 0.3 g of
solution; an orange-red precipitate is produced. Oxyphenbutazone with 60 ml of acetone, filter and evaporate
the filtrate to dryness. The residue complies with the following
Tests tests.

Appearance of solution. To 0.5 g add a mixture of 12 ml of 1 M A. When examined in the range 230 nm to 360 nm (2.4.7), a
sodium hydroxide and 8 ml of a 7.5 per cent w/v solution of 0.001 per cent w/v solution in 0.01 M sodium hydroxide shows
glycine, shake for 1 minute and maintain at 25º for exactly an absorption maximum at about 254 nm; absorbance at about
60 minutes. The solution is clear (2.4.1). 254 nm, 0.71 to 0.77.
B. To 2 ml of a 1 per cent w/v solution in ethanol (95 per cent)
Related substances. Determine by thin-layer chromatography
add 2 ml of a 0.1 per cent w/v solution of 2,6-dichloroquinone-
(2.4.17), coating the plate with silica gel HF254.
4-chlorimide in ethanol (95 per cent) and 1 ml of sodium
Mobile phase. A 0.02 per cent w/v solution of butylated carbonate solution; an intense green colour is produced.
hydroxytoluene in a mixture of 80 volumes of chloroform and
C. To 0.1 g add 1 ml of glacial acetic acid and 2 ml of
20 volumes of glacial acetic acid.
hydrochloric acid and boil under a reflux condenser for
Test solution. Dissolve 0.1 g of the substance under 30 minutes. Cool, add 10 ml of water and filter. To the filtrate
examination in 5 ml of a solution containing 0.02 per cent w/v add 3 ml of a 0.7 per cent w/v solution of sodium nitrite; a
of butylated hydroxytoluene in ethanol. yellow colour develops. To 1 ml of the resulting solution add
a solution of 10 mg of 2-naphthol in 5 ml of sodium carbonate
Reference solution. Dilute 1 ml of the test solution to 200 ml
solution; an orange-red precipitate is produced.
with the ethanolic butylated hydroxytoluene solution.
To prepare the plate, allow the mobile phase to rise 4 cm. Dry Tests
the plate in a current of cold air for 1 minute. Without delay Related substances. Determine by thin-layer chromatography
and under a current of nitrogen, apply separately to the plate (2.4.17), coating the plate with silica gel HF254.
5 µl of each solution prepared immediately before use. Develop
immediately, allowing the mobile phase to rise 10 cm. Dry the Mobile phase. A 0.02 per cent w/v solution of butylated
plate in a current of cold air for 15 minutes and examine in hydroxytoluene in a mixture of 80 volumes of chloroform and
ultraviolet light at 254 nm. Any secondary spot in the 20 volumes of glacial acetic acid.
chromatogram obtained with the test solution is not more Test solution. Shake a quantity of the powdered tablets
intense than the spot in the chromatogram obtained with the containing 0.2 g of Oxyphenbutazone with 10 ml of a solution
reference solution. containing 0.02 per cent w/v of butylated hydroxytoluene in
ethanol for 15 minutes and centrifuge. Use the supernatant
Heavy metals (2.3.13). 1.0 g complies with the limit test for
liquid.
heavy metals, Method B (20 ppm).
Reference solution. Dilute 1 ml of the test solution to 200 ml
Sulphated ash (2.3.18). Not more than 0.1 per cent.
with the ethanolic butylated hydroxytoluene solution.
Water (2.3.43). 5.0 to 6.0 per cent, determined on 0.5 g. To prepare the plate, allow the mobile phase to rise 4 cm. Dry
Assay. Weigh accurately about 0.5 g, dissolve in 50 ml of the plate in a current of cold air for 1 minute. Without delay
acetone and titrate with 0.1 M sodium hydroxide, using and under a current of nitrogen, apply separately to the plate
bromothymol blue solution as indicator and continuing the 5 µl of each solution prepared immediately before use. Develop
titration until the blue colour persists for at least 15 seconds. immediately, allowing the mobile phase to rise 10 cm. Dry the
Carry out a blank titration. plate in a current of cold air for 15 minutes and examine in
ultraviolet light at 254 nm. Any secondary spot in the
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03244 g of
chromatogram obtained with the test solution is not more
C19H20N2O3.
intense than the spot in the chromatogram obtained with the
Storage. Store protected from light. reference solution.

874
IP 2007 OXYTETRAXYCLINE DIHYDRATE

Dissolution (2.5.2). Oxytetracycline has a potency not less than 900 µg of


Apparatus. No 1 C22H24N2O9, per mg, calculated on the anhydrous basis.
Medium. 900 ml of phosphate buffer pH 7.6. Description. A tan yellow or light yellow (with or without a
Speed and time. 100 rpm and 30 minutes. greenish tinge), crystalline powder; odourless.
Withdraw a suitable volume of the medium and filter promptly Identification
through a membrane filter disc with an average pore diameter
not greater than 1.0 µm. Reject the first few ml of the filtrate A. Determine by thin-layer chromatography (2.4.17), coating
and dilute a suitable volume of the filtrate with the same the plate with the substance prepared by mixing 25 g of silica
solvent. Measure the absorbance of the resulting solution at gel G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of
the maximum at about 262 nm (2.4.7). Calculate the content of 0.1 M disodium edetate previously adjusted to pH 7.0 with
C19H20N2O3,H2O in the medium form the absorbance obtained dilute ammonia solution. After spreading the plate, allow it to
from a solution of known concentration of oxyphenbutazone stand at room temperature till it is dry (70 to 90 minutes).
RS in the same medium.
Mobile phase. The lower layer formed after shaking 200 ml of
D. Not less than 60 per cent of the stated amount of a mixture of 2 volumes of ethyl acetate, 2 volumes of
C19H20N2O3,H2O. chloroform and 1 volume of acetone with 25 ml of 0.1 M
Other Tests. Comply with the tests stated under Tablets. disodium edetate previously adjusted to pH 7.0 with dilute
ammonia solution.
Assay. Weigh 20 tablets and reduce to a fine powder. Weigh
accurately a quantity of the powder containing about 0.5 g of Test solution. Dissolve 0.05 g of the substance under
Oxyphenbutazone and extract successively with 30, 10 and examination in 100 ml of methanol.
10 ml of warm acetone. Filter the combined extracts, cool. Reference solution (a). A 0.05 per cent w/v solution of
Titrate with 0.1 M sodium hydroxide, using bromothymol oxytetracycline RS in methanol.
blue solution as indicator and continuing the titration until
Reference solution (b). A solution containing 0.05 per cent
the blue colour persists for at least 15 seconds. Carry out a
w/v each of demethylchlortetracycline hydrochloride RS,
blank titration.
oxytetracycline hydrochloride RS and tetracycline
1 ml of 0.1 M sodium hydroxide is equivalent to 0.03424 g of hydrochloride RS in methanol.
C19H20N2O3,H2O.
Apply to the plate 1 µl of each solution, freshly prepared.
Storage. Store protected from moisture. After development, dry the plate in air, expose to the vapours
of ammonia and examine in ultraviolet light at 365 nm. The
principal spot in the chromatogram obtained with the test
solution corresponds to that in the chromatogram obtained
Oxytetracycline Dihydrate with reference solution (a). The test is not valid unless the
chromatogram obtained with reference solution (b) shows
Oxytetracycline three clearly separated spots.
B. To about 0.5 mg add 2 ml of sulphuric acid; a red colour is
OH O OH O O produced. Add the solution to 1 ml of water; the colour
OH changes to yellow.
NH2
, 2H2O C. Dissolve about 2 mg in 5 ml of a 1 per cent w/v solution of
H OH sodium carbonate and add 2 ml of diazotised sulphanilic
H
H3C OH OH N acid solution; an orange-red to brownish-red colour is
H3C CH3 produced.

Tests
C22H24N2O9,2H2O Mol. Wt. 496.5
Oxytetracycline is (4S,4aR,5S,5aR,6S,12aS)-4-dimethyl pH (2.4.24). 4.5 to 7.5, determined in a 1.0 per cent w/v
amino-1,4,4a,5,5a,6,11,12a-octahydro-3,5,6,10,12,12a- suspension in freshly boiled and cooled water.
hexahydroxy-6-methyl-1,11-dioxonaphthacene-2- Specific optical rotation (2.4.22). –203º to –216º, determined
carboxamide, a substance produced by the growth of at 20º in a 1.0 per cent w/v solution in 0.1 M hydrochloric
certain strains of Streptomyces rimosus or obtained by any acid, after allowing the solution to stand protected from light
other means. It contains a variable quantity of water. for 30 minutes before measurement.

875
OXYTETRAXYCLINE DIHYDRATE IP 2007

Light absorption. Absorbance of a 0.002 per cent w/v solution Adjust the sensitivity so that the heights of the peaks in the
in buffer solution pH 2.0 at the maximum at about 353 nm, 0.58 chromatogram obtained with reference solution (d) are at least
to 0.62 (2.4.7). 50 per cent of the full-scale deflection of the recorder.
Light-absorbing impurities. A. Dissolve 20 mg in sufficient The test is not valid unless (a) the resolution between the first
of a mixture of 1 volume of 1 M hydrochloric acid and peak (4-epioxytetracycline) and the second peak
99 volumes of methanol to produce 10 ml. Absorbance of the (oxytetracycline) is at least 4.0 (b) the resolution between the
resulting solution at about 430 nm, when measured within second peak and the third peak (tetracycline) is at least 5.0
1 hour of preparing the solution, not more than 0.25, calculated (the content of 2-methyl-2-propanol in the mobile phase may
on the anhydrous basis (2.4.7). be adjusted if necessary) and (c) the symmetry factor for the
second peak is at most 1.25.
B. Dissolve 0.1 g in sufficient of a mixture of 1 volume of 1 M
hydrochloric acid and 99 volumes of methanol to produce Inject reference solution (a) six times. The test is not valid
10 ml. Absorbance of the resulting solution at about 490 nm, unless the relative standard deviation of the area of the peak
when measured within 1 hour of preparing the solution, not due to oxytetracycline is not greater than 1.0 per cent.
more than 0.20, calculated on the anhydrous basis (2.4.7).
Inject the test solution and reference solution (e). In the
Related substances. Determine by liquid chromatography chromatogram obtained with the test solution the area of any
(2.4.14). peak corresponding to 4-epioxytetracycline or tetracycline is
not greater than the area of the corresponding peak in the
Test solution. Dissolve 20 mg of the substance under
chromatogram obtained with reference solution (e). In the
examination in 25 ml of 0.01 M hydrochloric acid.
chromatogram obtained with the test solution the area of any
Reference solution (a). Dissolve 20 mg of oxytetracycline RS peak appearing on the tail of the principal peak is not greater
in 25 ml of 0.01 M hydrochloric acid. than 4.0 times that of the peak due to 4-epioxytetracycline in
the chromatogram obtained with reference solution (e).
Reference solution (b). Dissolve 20 mg of 4-epioxytetra-
cycline RS in 25 ml of 0.01 M hydrochloric acid. Heavy metals (2.3.13). 0.4 g complies with limit the test for
heavy metals, Method B (50 ppm).
Reference solution (c). Dissolve 20 mg of tetracycline
hydrochloride RS in 25 ml of 0.01 M hydrochloric acid. Sulphated ash (2.3.18). Not more than 0.5 per cent.
Reference solution (d). Dilute a mixture of 1.5 ml of reference Water (2.3.43). 4.0 to 9.0 per cent, determined on 0.5 g.
solution (a), 1.0 ml of reference solution (b) and 3.0 ml of
Assay. Determine by the microbiological assay of antibiotics,
reference solution (c) to 25 ml with 0.01 M hydrochloric
Method A or B (2.2.10), and express the results in µg of
acid.
oxytetracycline, C22H24N2O9, per mg.
Reference solution (e). Dilute a mixture of 1.0 ml of reference
Oxytetracycline intended for use in the manufacture of
solution (b) and 4.0 ml of reference solution (c) to 200 ml with
parenteral preparations without a further procedure for the
0.01 M hydrochloric acid.
removal of bacterial endotoxins complies with the following
Chromatographic system additional requirement.
– a stainless steel column 25 cm x 4.6 mm, packed with
Bacterial endotoxins (2.2.3). Not more than 0.4 Endotoxin Unit
styrene- divinylbenzene co-polymer (8 to 10 µm),
per mg of Oxytetracycline.
– column. temperature 60º,
– mobile phase: add 60 g of 2-methyl-2-propanol to a Oxytetracycline intended for use in the manufacture of
volumetric flask with the aid of 200 ml of water, add parenteral preparations without a further sterilisation
60 ml of 0.33 M phosphate buffer pH 7.5, 50 ml of a procedure complies with the following additional
1.0 per cent w/v solution of tetrabutylammonium requirement.
hydrogen sulphate previously adjusted to pH 7.5 with Sterility. Complies with the test for sterility (2.2.11).
2 M sodium hydroxide and 10 ml of a 0.04 per cent w/v
solution of disodium edetate previously adjusted to Storage. Store protected from light and moisture. If it is
pH 7.5 with 2 M sodium hydroxide and dilute to intended for use in the manufacture of parenteral preparations,
1000.0 ml with water, the container should be sterile and sealed so as to exclude
– flow rate. 1 ml per minute, micro-organisms.
– spectrophotometer set at 254 nm, Labelling. The label states whether or not the contents are
– a 20 µl loop injector. intended for use in the manufacture of parenteral preparations.

876
IP 2007 OXYTETRAXYCLINE HYDROCHLORIDE

Oxytetracycline Injection Other Tests. Complies with the tests stated under Parenteral
Preparations (Injections).
Oxytetracycline Injection is a sterile solution of oxytetracycline
Assay. Determine by the microbiological assay of antibiotics,
with or without one or more suitable buffering agents,
Method A or B (2.2.10), and express the result in mg of
anaesthetics, preservatives, antioxidants, complexing agents
oxytetracycline, C22H24N2O9, per ml.
and solvents.
Storage. Store protected from light.
Oxytetracycline Injection contains not less than 90.0 per cent
and not more than 120.0 per cent of the stated amount of Labelling. The label states (1) the strength in mg of anhydrous
anhydrous oxytetracycline, C22H24N2O9. oxytetracycline per ml; (2) that the contents are to be used for
intramuscular use only; (3) the names of any preservatives
Description. A clear, yellow to tan yellow solution. It may used.
have a greenish tinge.

Identification
Oxytetracycline Hydrochloride
A. Determine by thin-layer chromatography (2.4.17), coating
the plate with the substance prepared by mixing 25 g of silica C22H24N2O9,HCl Mol. Wt. 496.9
gel G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of Oxytetracycline Hydrochloride is (4S,4aR,5S,5aR,6S, 12aS)-
0.1 M disodium edetate previously adjusted to pH 7.0 with 4-dimethylamino-1,4,4a,5,5a,6,11,12a-octahydro-3,5,
dilute ammonia solution. After spreading the plate, allow it to 6,10,12,12a-hexahydroxy-6-methyl-1,11-dioxonaphthacene-2-
stand at room temperature till it is dry (70 to 90 minutes). carboxamide hydrochloride, a substance produced by
Mobile phase. The lower layer formed after shaking 200 ml of the growth of certain strains of Streptomyces rimosus or
a mixture of 2 volumes of ethyl acetate, 2 volumes of obtained by any other means.
chloroform and 1 volume of acetone with 25 ml of 0.1 M Description. A pale yellow, crystalline powder; odourless;
disodium edetate previously adjusted to pH 7.0 with dilute hygroscopic.
ammonia solution.
Oxytetracycline Hydrochloride has a potency not less than
Test solution. Shake a quantity containing 10 mg of anhydrous 835 µg of C22H24N2O9, per mg, calculated on the anhydrous
oxytetracycline with 20 ml of methanol, centrifuging if basis.
necessary and use the clear supernatant liquid.
Reference solution (a). A 0.05 per cent w/v solution of Identification
oxytetracycline hydrochloride RS in methanol. A. Determine by thin-layer chromatography (2.4.17), coating
Reference solution (b). A solution containing 0.05 per cent the plate with the substance prepared by mixing 25 g of silica
w/v each of demethylchlortetracycline hydrochloride RS, gel G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of
oxytetracycline hydrochloride RS and tetracycline 0.1 M disodium edetate previously adjusted to pH 7.0 with
hydrochloride RS in methanol. dilute ammonia solution. After spreading the plate, allow it to
stand at room temperature till it is dry (70 to 90 minutes).
Apply to the plate 1 µl of each solution, freshly prepared.
After development, dry the plate in air, expose to the vapours Mobile phase. The lower layer formed after shaking 200 ml of
of ammonia and examine in ultraviolet light at 365 nm. The a mixture of 2 volumes of ethyl acetate, 2 volumes of
principal spot in the chromatogram obtained with the test chloroform and 1 volume of acetone with 25 ml of 0.1 M
solution corresponds to that in the chromatogram obtained disodium edetate previously adjusted to pH 7.0 with dilute
with reference solution (a). The test is not valid unless the ammonia solution.
chromatogram obtained with reference solution (b) shows Test solution. Dissolve 0.05 g of the substance under
three clearly separated spots. examination in 100 ml of methanol.
B. Add 0.1 ml to 2 ml of sulphuric acid; a red colour is produced. Reference solution (a). A 0.05 per cent w/v solution of
Add the solution to 1 ml of water; the colour changes to oxytetracycline hydrochloride RS in methanol.
yellow.
Reference solution (b). A solution containing 0.05 per cent
Tests w/v each of demethylchlortetracycline hydrochloride RS,
oxytetracycline hydrochloride RS and tetracycline
pH (2.4.24). 8.0 to 9.0. hydrochloride RS in methanol.
Bacterial endotoxins (2.2.3). Not more than 0.4 Endotoxin Unit Apply to the plate 1 µl of each solution, freshly prepared.
per mg of Oxytetracycline. After development, dry the plate in air, expose to the vapours

877
OXYTETRAXYCLINE HYDROCHLORIDE IP 2007

of ammonia and examine in ultraviolet light at 365 nm. The Reference solution (e). Dissolve 20 mg of b -apo-
principal spot in the chromatogram obtained with the test oxytetracycline RS in 20 ml of 0.01 M sodium hydroxide and
solution corresponds to that in the chromatogram obtained dilute to 250 ml with 0.01 M hydrochloric acid.
with reference solution (a). The test is not valid unless the
Reference solution (f). Dilute a mixture of 1.5 ml of reference
chromatogram obtained with reference solution (b) shows
solution (a), 1.0 ml of reference solution (b), 3.0 ml each of
three clearly separated spots.
reference solutions (c) (d) and (e) to 25 ml with 0.01 M
B. To about 0.5 mg add 2 ml of sulphuric acid; a red colour is hydrochloric acid.
produced. Add the solution to 1 ml of water; the colour
Reference solution (g). Dilute a mixture of 1.0 ml of reference
changes to yellow.
solution (b), 4.0 ml of reference solution (c) and 40.0 ml of
C. Dissolve about 2 mg in 5 ml of a 1 per cent w/v solution of reference solution (e) to 200 ml with 0.01 M hydrochloric
sodium carbonate and add 2 ml of diazotised sulphanilic acid.
acid solution; an orange-red to brownish-red colour is
Chromatographic system
produced.
– a stainless steel column 25 cm x 4.6 mm, packed with
D. A 5 per cent w/v solution gives the reactions of chlorides styrene-divinylbenzene co-polymer (8 to 10 µm),
(2.3.1). – column. temperature 60º,
– mobile phase: transfer separately 30 g (for mobile phase
Tests A) or 100 g (for mobile phase B) of 2-methyl-2-propanol
pH (2.4.24). 2.0 to 3.0, determined in a 1.0 per cent w/v solution. to volumetric flasks with the aid of 200 ml of water; to
each flask add 60 ml of 0.33 M phosphate buffer pH 7.5,
Specific optical rotation (2.4.22). –188º to –200º, determined 50 ml of a 1.0 per cent w/v solution of tetrabutyl-
at 20º in a 1 per cent w/v solution in 0.1 M hydrochloric acid. ammonium hydrogen sulphate previously adjusted to
Light absorption (2.4.7). Absorbance of a 0.002 per cent w/v pH 7.5 with 2 M sodium hydroxide and 10 ml of a 0.04
solution in chloride buffer solution pH 2.0 at the maximum at per cent w/v solution of disodium edetate previously
about 353 nm, 0.54 to 0.58. adjusted to pH 7.5 with 2 M sodium hydroxide and
dilute each solution to 1000.0 ml with water.,
Light-absorbing impurities. A. Dissolve 20 mg in sufficient
– flow rate. 1 ml per minute,
of a mixture of 1 volume of 1 M hydrochloric acid and
– spectrophotometer set at 254 nm,
99 volumes of methanol to produce 10 ml. Absorbance of the
resulting solution at about 430 nm, when measured within – a 20 µl loop injector.
1 hour of preparing the solution, not more than 0.50, calculated Carry out a one-step gradient elution in the following manner.
on the anhydrous basis (2.4.7). Pump a mixture containing 30 volumes of mobile phase B and
B. Dissolve 0.1 g in sufficient of a mixture of 1 volume of 1 M 70 volumes of mobile phase A for 15 minutes, then pump a
hydrochloric acid and 99 volumes of methanol to produce mixture containing 30 volumes of mobile phase A and
10 ml. Absorbance of the resulting solution at about 490 nm, 70 volumes of mobile phase B for 15 minutes and finally
when measured within 1 hour of preparing the solution, not equilibrate with the first mixture. Adjust the sensitivity so that
more than 0.20, calculated on the anhydrous basis (2.4.7). the heights of the peaks in the chromatogram obtained with
reference solution (f) are at least 50 per cent of full-scale
Related substances. Determine by liquid chromatography deflection of the recorder.
(2.4.14).
The test is not valid unless, in the chromatogram obtained
Test solution. Dissolve 20 mg of the substance under with reference solution (f), (a) the resolution between the first
examination in 25 ml of 0.01 M hydrochloric acid. peak (4-epioxytetracycline) and the second peak
Reference solution (a). Dissolve 20 mg of oxytetracycline RS (oxytetracycline) is at least 4.0, (b) the resolution between the
in 25 ml of 0.01 M hydrochloric acid. second peak and the third peak (tetracycline) is at least 5.0, (c)
the resolution between the fourth peak (a -apo-oxytetracycline)
Reference solution (b). Dissolve 20 mg of 4-epioxytetra- and the fifth peak (b-apo-oxytetracycline) is at least 3.5, and
cycline RS in 25 ml of 0.01 M hydrochloric acid. (d) the symmetry factor of the second peak is at most 1.25. If
Reference solution (c). Dissolve 20 mg of tetracycline necessary adjust the proportions of the mobile phases used
hydrochloride RS in 25 ml of 0.01 M hydrochloric acid. to produce the one-step gradient elution. Adjust the time-
programme for the one-step gradient elution if necessary.
Reference solution (d). Dissolve 20 mg of a-apo-
oxytetracycline RS in 20 ml of 0.01 M sodium hydroxide and Inject reference solution (a) six times. The test is not valid if
dilute to 250 ml with 0.01 M hydrochloric acid. the relative standard deviation of the area of the peak due to

878
IP 2007 OXYTETRAXYCLINE CAPSULES

oxytetracycline is greater than 1.0 per cent. If necessary, adjust Identification


the integrator parameters.
A. Determine by thin-layer chromatography (2.4.17), coating
Inject the test solution and reference solution (g). In the the plate with the substance prepared by mixing 25 g of silica
chromatogram obtained with the test solution the area of any gel G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of
peak corresponding to 4-epioxytetracycline or tetracycline 0.1 M disodium edetate previously adjusted to pH 7.0 with
is not greater than the area of the corresponding peak in the dilute ammonia solution. After spreading the plate, allow it to
chromatogram obtained with reference solution (g) and the stand at room temperature till it is dry (70 to 90 minutes).
total area of the peaks corresponding to a -apo-oxytetracycline
and to b -apo-oxytetracycline and any peak between the latter Mobile phase. The lower layer formed after shaking 200 ml of
two is not greater than the area of the peak due to b -apo- a mixture of 2 volumes of ethyl acetate, 2 volumes of
oxytetracycline in the chromatogram obtained with reference chloroform and 1 volume of acetone with 25 ml of 0.1 M
solution (g). In the chromatogram obtained with the test disodium edetate previously adjusted to pH 7.0 with dilute
solution the area of any peak appearing on the tail of the ammonia solution.
principal peak is not greater than 4.0 times that of the peak due Test solution. Extract a quantity of the contents of the capsules
to 4-epioxytetracycline in the chromatogram obtained with containing 10 mg of Oxytetracycline Hydrochloride with 20 ml
reference solution (g). methanol, centrifuge and use the supernatant liquid.
Heavy metals (2.3.13). 0.4 g complies with the limit test for Reference solution (a). A 0.05 per cent w/v solution of
heavy metals, Method B (50 ppm). oxytetracycline hydrochloride RS in methanol.
Sulphated ash (2.3.18). Not more than 0.5 per cent.
Reference solution (b). A solution containing 0.05 per cent
Water (2.3.43). Not more than 2.0 per cent w/w, determined on w/v each of demethylchlortetracycline hydrochloride RS,
0.5 g. oxytetracycline hydrochloride RS and tetracycline
Assay. Determine by the microbiological assay of antibiotics, hydrochloride RS in methanol.
Method A or B (2.2.10), and express the result in µg of Apply to the plate 1 µl of each solution, freshly prepared.
oxytetracycline, C22H24N2O9, per mg. After development, dry the plate in air, expose to the vapours
Oxytetracycline Hydrochloride intended for use in the of ammonia and examine in ultraviolet light at 365 nm. The
manufacture of parenteral preparations without a further principal spot in the chromatogram obtained with the test
procedure for the removal of bacterial endotoxins complies solution corresponds to that in the chromatogram obtained
with the following additional requirement. with reference solution (a). The test is not valid unless the
Bacterial endotoxins (2.2.3). Not more than 0.4 Endotoxin Unit chromatogram obtained with reference solution (b) shows
per mg. three clearly separated spots.

Oxytetracycline Hydrochloride intended for use in the B. To 0.5 mg of the contents of the capsules add 2 ml of
manufacture of parenteral preparations without a further sulphuric acid; a red colour is produced. Add the solution to
sterilisation procedure complies with the following 1 ml of water; the colour changes to yellow.
additional requirement. C. Dissolve about 2 mg of the contents of the capsules in 5 ml
Sterility. Complies with test for sterility (2.2.11). of a 1 per cent w/v solution of sodium carbonate and add 2 ml
of diazotised sulphanilic acid solution; a light brown colour
Storage. Store protected from light and moisture. If it is
is produced.
intended for use in the manufacture of parenteral preparations,
the container should be sterile and sealed so as to exclude D. The contents of the capsules give the reactions of chlorides
micro-organisms. (2.3.1).
Labelling. The label states whether or not the contents are
intended for use in the manufacture of parenteral preparations. Tests
Light-absorbing impurities. Dissolve a portion of the mixed
contents of five capsules as completely as possible in
Oxytetracycline Capsules sufficient of a mixture of 1 volume of 1 M hydrochloric acid
and 99 volumes of methanol to produce two solutions of
Oxytetracycline Hydrochloride Capsules Oxytetracycline Hydrochloride containing (1) 0.2 per cent w/v
Oxytetracycline Capsules contain not less than 95.0 per cent and (2) 1.0 per cent w/v and filter each solution. Absorbance
and not more than 110.0 per cent of the stated amount of of the filtrate obtained from solution (1) at about 430 nm, when
oxytetracycline hydrochloride, C22H24N2O9,HCl. measured within 1 hour of preparing the solution, not greater

879
OXYTETRAXYCLINE EYE OINTMENT IP 2007

than 0.75 and of the filtrate obtained from solution (2) at about Test solution. A solution prepared by heating a quantity
490 nm, not more than 0.40 (2.4.7). containing 20 mg of Oxytetracycline Hydrochloride with 20 ml
Dissolution (2.5.2). of methanol for 20 minutes, cooling in ice, filtering, carefully
evaporating the filtrate to dryness and dissolving the residue
Apparatus. No 1 in 20 ml of methanol.
Medium. 900 ml of 0.1 M hydrochloric acid.
Reference solution (a). A 0.05 per cent w/v solution of
Speed and time. 100 rpm and 45 minutes. oxytetracycline hydrochloride RS in methanol.
Withdraw a suitable volume of the medium and filter through
Reference solution (b). A solution containing 0.05 per cent
a membrane filter disc with an average pore diameter not greater
w/v each of demethylchlortetracycline hydrochloride RS,
than 1.0 µm. Measure the absorbance of the filtrate, suitably
oxytetracycline hydrochloride RS and tetracycline
diluted if necessary, at the maximum at about 353 nm (2.4.7).
hydrochloride RS in methanol.
Calculate the content of C22H24N2O9,HCl in the medium taking
282 as the specific absorbance at 353 nm. Apply to the plate 1 µl of each solution, freshly prepared.
After development, dry the plate in air, expose to the vapours
D. Not less than 75 per cent of the stated amount of
of ammonia and examine in ultraviolet light at 365 nm. The
C22H24N2O9,HCl.
principal spot in the chromatogram obtained with the test
Loss on drying (2.4.19). Not more than 5.0 per cent, determined solution corresponds to that in the chromatogram obtained
on 1.0 g of the mixed contents of the capsules by drying in an with reference solution (a). The test is not valid unless the
oven at 60º at a pressure not exceeding 0.7 kPa for 3 hours. chromatogram obtained with reference solution (b) shows
Other Tests. Comply with the tests stated under Capsules. three clearly separated spots.

Assay. Weigh accurately a quantity of the mixed contents of Tests


20 capsules containing about 0.25 g of Oxytetracycline
Hydrochloride, add 250.0 ml of water, shake, filter. Water (2.3.43). Not more than 1.0 per cent, determined on 0.5 g.

Determine by the microbiological assay of antibiotics, Method Other Tests. Complies with the tests stated under Eye
A or B (2.2.10), and express the results in mg of oxytetracycline Ointments.
hydrochloride per capsule taking each mg of oxytetracycline Assay. Weigh accurately about 1.0 g and transfer to a
to be equivalent to 1.079 mg of oxytetracycline hydrochloride, separating funnel. Add 25 ml of peroxide-free ether, shake
C22H24N2O9,HCl. well and extract with five quantities, each of 20 ml, of 0.1 M
Storage. Store protected from light and moisture. hydrochloric acid. Combine the extracts and dilute to
200.0 ml with 0.1 M hydrochloric acid. Dilute a suitable volume
of the resulting solution with buffer solution No 3 (2.2.10), to
produce a solution containing 1 µg of oxytetracycline per ml.
Oxytetracycline Eye Ointment
Determine by the microbiological assay of antibiotics, Method
Oxytetracycline Hydrochloride Eye Ointment B (2.2.10), and express the results as a percentage of
Oxytetracycline Eye Ointment contains not less than 90.0 per oxytetracycline hydrochloride taking each mg of
cent and not more than 115.0 per cent of the stated amount of oxytetracycline to be equivalent to 1.079 mg of oxytetracycline
oxytetracycline hydrochloride, C22H24N2O9,HCl. hydrochloride, C22H24N2O9,HCl.
Storage. Store protected from light and moisture.
Identification
Determine by thin-layer chromatography (2.4.17), coating the
plate with the substance prepared by mixing 25 g of silica gel Oxytetracycline Hydrochloride
G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of
0.1 M disodium edetate previously adjusted to pH 7.0 with
Injection
dilute ammonia solution. After spreading the plate, allow it to Oxytetracycline Hydrochloride Injection is a sterile material
stand at room temperature till it is dry (70 to 90 minutes). consisting of Oxytetracycline Hydrochloride with or without
Mobile phase. The lower layer formed after shaking 200 ml of buffering agents and other excipients. It is filled in a sealed
a mixture of 2 volumes of ethyl acetate, 2 volumes of container.
chloroform and 1 volume of acetone with 25 ml of 0.1 M The injection is constituted by dissolving the contents of the
disodium edetate previously adjusted to pH 7.0 with dilute sealed container in the requisite amount of sterile Water for
ammonia solution. Injections, immediately before use.

880
IP 2007 OXYTOCIN

The constituted solution complies with the requirements for Tests


Clarity of solution and Particulate matter stated under
Parenteral Preparations (Injections). Appearance of solution. A 10.0 per cent w/v solution is clear
(2.4.1) and yellow.
Storage. The constituted solution may be used within three
days of preparation when stored in a refrigerator (2º to 8º). pH (2.4.24). 2.0 to 3.0, determined in a 1.0 per cent w/v solution.

Oxytetracycline Hydrochloride Injection contains not less than Light-absorbing impurities. A. Dissolve 20 mg in sufficient
90.0 per cent and not more than 115.0 per cent of the stated of a mixture of 1 volume of 1 M hydrochloric acid and
amount of oxytetracycline, C22H24N2O9. 99 volumes of methanol to produce 10 ml. Absorbance of the
resulting solution at about 430 nm, when measured within
The contents of the sealed container comply with the 1 hour of preparing the solution, not more than 0.50, calculated
requirements stated under Parenteral Preparations on the anhydrous basis (2.4.7).
(Powders for Injection) and with the following requirements.
B. Dissolve 0.1 g in sufficient of a mixture of 1 volume of 1 M
Description. A pale yellow, crystalline powder. hydrochloric acid and 99 volumes of methanol to produce
10 ml. Absorbance of the resulting solution at about 490 nm,
Identification when measured within 1 hour of preparing the solution, not
A. Determine by thin-layer chromatography (2.4.17), coating more than 0.20, calculated on the anhydrous basis (2.4.7).
the plate with the substance prepared by mixing 25 g of silica Assay. Determine by the microbiological assay of antibiotics,
gel G with 50 ml of a mixture of 2.5 ml of glycerin and 47.5 ml of method A or B (2.2.10), and express the result in µg of
0.1 M disodium edetate previously adjusted to pH 7.0 with oxytetracycline, C22H24N2O9, per mg.
dilute ammonia solution. After spreading the plate, allow it to
Bacterial endotoxins (2.2.3). Not more than 0.4 Endotoxin Unit
stand at room temperature till it is dry (70 to 90 minutes).
per mg.
Mobile phase. The lower layer formed after shaking 200 ml of
Storage. Store protected from light and moisture.
a mixture of 2 volumes of ethyl acetate, 2 volumes of
chloroform and 1 volume of acetone with 25 ml of 0.1 M Labelling. The label states (1) the quantity of Oxytetracycline
disodium edetate previously adjusted to pH 7.0 with dilute Hydrochloride contained in it in terms of the equivalent
ammonia solution. amount of oxytetracycline; (2) that the contents are to be
used for intravenous injection only; (3) the names of the
Test solution. Dissolve 0.05 g of the substance under buffering agents used.
examination in 100 ml of methanol.
Reference solution (a). A 0.05 per cent w/v solution of
oxytetracycline hydrochloride RS in methanol. Oxytocin
Reference solution (b). A solution containing 0.05 per cent
Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-GlyNH2
w/v each of demethylchlortetracycline hydrochloride RS,
oxytetracycline hydrochloride RS and tetracycline
hydrochloride RS in methanol.
C43H66N12O12S2 Mol. Wt.1007.2
Apply to the plate 1 µl of each solution, freshly prepared.
Oxytocin is a cyclic nonapeptide hormone obtained by a
After development, dry the plate in air, expose to the vapours
process of fractionation from the posterior lobe of the pituitary
of ammonia and examine in ultraviolet light at 365 nm. The
gland of healthy oxen or other mammals or by synthesis that
principal spot in the chromatogram obtained with the test
has the property of stimulating contraction of the uterus and
solution corresponds to that in the chromatogram obtained
milk ejection in receptive animals. It may be presented as a
with reference solution (a). The test is not valid unless the
solid or as a solution in a solvent containing an appropriate
chromatogram obtained with reference solution (b) shows
antimicrobial preservative such as 0.2 per cent w/v solution of
three clearly separated spots.
chlorbutol.
B. To about 0.5 mg add 2 ml of sulphuric acid; a red colour is
If it is derived from animal species, Oxytocin contains not less
produced. Add the solution to 1 ml of water; the colour
than 90.0 per cent and not more than 111.0 per cent of the
changes to yellow.
stated number of Units of oxytocic activity. If it is a synthetic
C. Dissolve about 2 mg in 5 ml of a 1 per cent w/v solution of product presented as a solid, it contains not less than
sodium carbonate and add 2 ml of diazotised sulphanilic 560 Units per mg, calculated with reference to the peptide
acid solution; an orange-red to brownish-red colour is content and when presented as a liquid, it contains not less
produced. than 150 Units per ml.

881
OXYTOCIN IP 2007

Description. When presented as a solid, a white or almost Method A. By depression of the blood pressure in chicken —
white powder. When presented as a liquid, a clear colourless Anaesthetise a young healthy adult cockerel weighing 1.2 to
liquid. 2.3 kg with an anaesthetic that will maintain a prolonged and
constant high blood pressure. Expose the gluteus primus
Identification muscle in one thigh and cut and retract it to reveal the popliteal
Gives rise to an appropriate response when administered as artery and crural vein. Cannulate the popliteal artery and record
directed under one of the methods described for Assay. the blood pressure on a suitable recorder calibrated for use
over a linear range. Cannulate the crural or brachial vein.
Tests
Immediately before use prepare a solution of the Standard
Peptide. 90.0 to 110.0 per cent of the stated amount of oxytocin, Preparation in saline solution so that the volume to be injected
C43H66N12O12S2 expressed per mg for the solid, and in mg per is between 0.1 ml and 0.5 ml. Record the blood pressure
ml for the liquid, responses to the injection into the cannulated vein of two
Determine by liquid chromatography (2.4.14). doses of this solution; the doses should be such as to produce
clearly discriminated, precipitous, submaximal decreases in
Test solution. Dissolve 3.5 mg of the substance under blood pressure; the required doses normally lie between
examination in sufficient of a 1.56 per cent w/v solution of 20 and 100 milliUnits. The interval between injections should
sodium dihydrogen phosphate to produce 10.0 ml or use the be constant and lie between 3 and 10 minutes depending on
liquid preparation as appropriate. the rate at which the blood pressure returns to normal.
Reference solution. Dissolve 3.5 mg of oxytocin RS in sufficient Immediately before use dilute the preparation being examined
of a 1.56 per cent w/v solution of sodium dihydrogen with saline solution so as to obtain responses similar to those
phosphate to produce 10.0 ml. obtained with the Standard Preparation. The ratio between
Chromatographic system the two doses of the preparation under examination should be
– a stainless steel column 12 cm x 4.6 mm, packed with the same as that between the two doses of the Standard
octadecylsilane bonded to porous silica (5 µm), Preparation and this ratio should be kept constant throughout
– mobile phase: appropriate proportions of a 1.56 per cent the assay.
w/v solution of sodium dihydrogen phosphate (mobile The two doses of the Standard Preparation and the two doses
phase A) and a mixture of equal volumes of acetonitrile of the preparation under examination should be given
and water (mobile phase B), according to a randomised block or a Latin square design and
– flow rate. 1 ml per minute, at least six responses to each should be recorded.
– spectrophotometer set at 220 nm,
If the animal rapidly becomes insensitive to the repeated
– a 20 µl loop injector.
injections of the solutions another animal must be used.
Equilibrate the column with a mixture of 70 volumes of mobile Measure all the responses and calculate the result of the assay
phase A and 30 volumes of mobile phase B and record the by standard statistical methods.
chromatograms as follows. Operate by gradient elution
increasing continuously and linearly the proportion of mobile Method B. By contraction of the rat uterus — Inject 100 µg of
phase B by 1.0 per cent v/v per minute for 30 minutes. Finally oestradiol benzoate intramuscularly into a female rat weighing
elute using the same mixture for 15 minutes to re-equilibrate 120 to 200 g 18 to 24 hours before the assay. Immediately
the column. before the assay confirm by vaginal smear that the rat is in
oestrus or preoestrus. Kill the rat and suspend one horn of
Calculate the content of the peptide, C43H66N12O12S2. the uterus in a bath containing a solution of the following
Assay.The potency of oxytocin is determined by comparing composition.
its activity with that of the Standard Preparation of oxytocin Composition (per cent w/v)
under the conditions of a suitable method of assay.
Sodium chloride 0.662
Standard Preparation Potassium chloride 0.045
The Standard Preparation is the 4th International Standard for Calcium chloride 0.007
Oxytocin, established in 1978, consisting of freeze-dried Sodium bicarbonate 0.256
synthetic oxytocin peptide with human albumin and citric acid
Disodium hydrogen phosphate 0.029
(supplied in ampoules containing 12.5 Units), or any other
suitable preparation the potency of which has been determined Sodium dihydrogen phosphate 0.003
in relation to the International Standard. Magnesium chloride 0.010
NOTE — Any of the following methods may be followed. Dextrose 0.050

882
IP 2007 OXYTOCIN

Maintain the bath at a temperature of 32º or at some other the whole system with a 3.8 per cent w/v solution of sodium
suitable temperature at which spontaneous contractions of citrate or saline solution containing 50 Units of heparin
the uterus are abolished and the preparation maintains its sodium per ml to prevent clotting of milk. After cannulation,
sensitivity. Oxygenate the solution with a mixture of 95 per inject a small volume (0.05 to 0.2 ml) of this solution into the
cent of oxygen and 5 per cent of carbon dioxide and record teat duct through the transducer to clear the milk from the tip
the contractions of the muscle using a suitable instrument of the cannula. (This procedure may be repeated during the
giving a linear response (for example an isotonic lever with a assay should obstruction arise from milk ejected into the
load not exceeding 2 g). Record the contractions produced by cannula). Clamp the strain gauge so that a slight tension is
the addition to the bath of two doses of the Standard applied to the teat and its natural alignment is preserved and
Preparation suitably diluted with the above solution. The doses connect the gauge to a potentiometric recorder adjusted to
should be such as to produce clearly discriminated, give full-scale deflection for an increase in milk-ejection
submaximal contractions; the required doses normally lie pressure of about 5.3 kPa. Inject all solutions through the
between 10 and 50 micro Units per ml of bath liquid. When venous cannula using a 1-ml syringe graduated in 0.01 ml and
maximal contraction has been reached, replace the bath liquid wash them in with 0.2 ml of saline solution.
by a fresh solution. The doses should be added at regular Prepare a solution of the Standard Preparation and a solution
intervals of 3 to 5 minutes depending upon the rate of recovery of the preparation under examination in saline solution so
of the muscle. Dilute the preparation under examination so as that the volume to be injected is between 0.1 ml and 0.4 ml.
to obtain responses on the addition of two doses similar to Choose two doses of the Standard Preparation such that the
those obtained with the Standard Preparation. The ratio increase in milk-ejection pressure is about 1.35 kPa for the
between the two doses of the preparation under examination lower dose and about 2.7 kPa for the higher dose. As an initial
should be the same as that between the two doses of the approximation, a lower dose of between 0.1 and 0.4 milliUnit
Standard Preparation and this ratio should be kept constant and an upper dose of 1.5 to 2 times this amount may be tried.
throughout the assay. Choose two doses of the preparation under examination with
The two doses of Standard Preparation and the two doses of the same inter-dose ratio, matching the effects of the doses of
the preparation under examination should be given according the Standard Preparation as closely as possible. Inject the
to a randomised block or a Latin square design and at least six four doses (two doses of the Standard Preparation and two
responses to each should be recorded. doses of the preparation being examined) at intervals of 3 to
5 minutes. The two doses of Standard Preparation and the
Measure all the responses and calculate the result of the assay two doses of the preparation under examination should be
by standard statistical methods. given according to a randomised block or a Latin square design
Method C. By measurement of milk-ejection pressure in a and at least four responses to each should be recorded.
lactating rat— Select a lactating rat, in the third to twenty- Measure all the responses and calculate the result of the assay
first day after parturition and weighing about 300 g, separate by standard statistical methods.
it from the litter and 30 to 60 minutes later anaesthetise (for
The estimated potency is not less than 90 per cent and not
example, by the intraperitoneal injection of a solution of
more than 111 per cent of the stated potency. The fiducial
Pentobarbitone Sodium). Tie the rat to an operating table,
limits of error are not less than 80 per cent and not more than
maintained at 37º, by its hind legs leaving the front legs free.
125 per cent of the stated potency.
Cannulate the trachea with a short polyethylene tube of
internal diameter about 2.5 mm in such a manner so as to Oxytocin of natural origin obtained by extraction and
ensure a free airway; apply artificial respiration only if purification complies with the following additional
necessary. Cannulate an external jugular or femoral vein with requirement.
a polyethylene tube of internal diameter about 0.4 mm which Vasopressor activity. Not more than 0.5 Unit per 20 Units of
is filled with saline solution and closed with a pin. oxytocic activity, when assayed by the biological assay for
Shave the skin surrounding the inguinal and abdominal teats vasopressor activity described below.
and excise the tip of one teat, preferably the lower inguinal The vasopressor activity is estimated by comparing the activity
teat. Insert a polyethylene tube of internal diameter about 0.3 of the preparation under examination with that of the Standard
mm and external diameter about 0.6 mm, to a depth sufficient Preparation of arginine vasopressin under the conditions of a
to obtain appropriate measurement of pressure (3 to 10 mm suitable method of assay.
depth), into the primary teat duct which opens onto the cut
surface and tie firmly in place with a ligature. Connect this Standard Preparation
cannula with a suitable strain gauge transducer (such as that The Standard Preparation is the Ist International Standard for
used for recording arterial blood pressure in the rat) and fill Arginine vasopressin, established in 1978, consisting of

883
OXYTOCIN INJECTION IP 2007

freeze-dried synthetic arginine vasopressin peptide acetate ratio, matching the effects of the dose of the Standard
with human albumin and citric acid (supplied in ampoules Preparation as closely as possible. Inject doses at intervals of
containing 8.20 Units), or another suitable preparation the 10 to 15 minutes.
potency of which has been determined in relation to that of The two doses of the Standard Preparation and the two doses
the International Standard. of the preparation under examination should be given in a
Inject slowly into the tail vein of a male albino rat weighing randomised block or a Latin square design and four to five
about 300 g a solution of a suitable a-adrenoceptor blocking responses to each should be recorded.
agent, for example 10 ml per kg of body weight of a solution Measure all the responses and calculate the result of the assay
prepared by dissolving 5 mg of phenoxybenzamine by standard statistical methods.
hydrochloride in 0.1 ml of ethanol (95 per cent), adding
0.05 ml of 1 M hydrochloric acid and diluting to 5 ml with Oxytocin intended for use in the manufacture of parenteral
saline solution. After 18 hours, anaesthetise the rat with an preparations without a further procedure for the removal of
anaesthetic that will maintain a prolonged and uniform blood bacterial endotoxins.complies with the following additional
pressure. After 45 to 60 minutes, tie the rat on its back to the requirement.
operating table by its hind legs. Cannulate the trachea with a Bacterial endotoxins (2.2.3). Not more than 100 Endotoxin
short polyethylene tube of external diameter about 2.5 mm Units per 200 Units of oxytocin.
and dissect a carotid artery ready for cannulation. Then
Oxytocin intended for use in the manufacture of parenteral
cannulate the femoral vein close to the inguinal ligament.
preparations without a further sterilisation procedure
Retract the abdominal muscles to expose the inguinal ligament.
complies with the following additional requirement.
Retract the superficial pudendal vein to one side and dissect
the femoral vein towards the inguinal ligament from the Sterility (2.2.11). Complies with the test for sterility.
corresponding artery. When dissecting, a deep branch Storage. Store protected from moisture. If the substance is
reaching the femoral vein must be found and tied off to prevent intended for use in the manufacture of parenteral preparations,
bleeding during cannulation. Tie a short polyethylene cannula the container should be sterile, tamper-evident and sealed so
of external diameter about 1 mm into the femoral vein by two as to exclude micro-organisms.
ligatures and join by a short piece of flexible tubing to a 1-ml
burette with an attached thistle funnel containing saline Labelling. The label states (1) the number of Units of oxytocic
solution at about 37º. Firmly fix a wet absorbent cotton swab activity per mg (for solid) or per ml (for liquid); (2) either the
to the thigh so as to cover the incision and cannula. At this animal species from which it is obtained or whether it is
stage inject through the venous cannula 200 Units of heparin, synthetic, as appropriate; (3) whether or not the contents are
dissolved in saline solution, per 100 g of body weight. Then intended for use in the manufacture of parenteral preparations.
tie in a carotid cannula of external diameter about 1 mm and
connect by a column of saline solution containing heparin
with a suitable pressure measuring device such as a mercury
manometer of internal diameter about 2 to 3 mm. Oxytocin Injection
The central and peripheral nervous system including both Oxytocin Injection is a sterile solution of Oxytocin in Water
vagus and associated sympathetic nerves is left intact. No for Injections.
artificial respiration is necessary. Taking care that no air is Oxytocin Injection contains not less than 90.0 per cent and
injected, inject all solutions through the venous cannula by not more than 110.0 per cent of the stated number of Units of
means of a 1-ml syringe graduated in 0.01 ml and wash in with oxytocin activity.
0.2 ml of saline solution from the burette.
Description. A clear, colourless liquid.
Dilute the extract of the Standard Preparation and the
preparation under examination with saline solution so that Identification
the volume to be injected is between 0.1 ml and 0.5 ml.
Gives rise to an appropriate response when administered as
Choose two doses of the Standard Preparation such that the directed under one of the methods described for Assay.
elevation of the blood pressure is about 4 kPa for the lower
dose and about 7 kPa but always submaximal for the higher Tests
dose, the ratio of low to high dose being determined by the
pH (2.4.24). 3.5 to 4.5.
response and usually being 3 to 5. As an initial approximation
doses of 3 and 5 milliUnits may be tried. Choose two doses of Other Tests. Complies with the tests stated under Parenteral
the preparation under examination with the same inter-dose Preparations (Injections).

884
IP 2007 OXYTOCIN INJECTION

Bacterial endotoxins (2.2.3). Not more than 100 Endotoxin oestrus or preoestrus. Kill the rat and suspend one horn of
Units per 200 Units of oxytocin. the uterus in a bath containing a solution of the following
Assay.The potency of oxytocin is determined by comparing composition.
its activity with that of the Standard Preparation of oxytocin Composition (per cent w/v)
under the conditions of a suitable method of assay. Sodium chloride 0.662
Standard Preparation Potassium chloride 0.045
Calcium chloride 0.007
The Standard Preparation is the 4th International Standard for
Oxytocin, established in 1978, consisting of freeze-dried Sodium bicarbonate 0.256
synthetic oxytocin peptide with human albumin and citric acid Disodium hydrogen phosphate 0.029
(supplied in ampoules containing 12.5 Units), or any other Sodium dihydrogen phosphate 0.003
suitable preparation the potency of which has been determined Magnesium chloride 0.010
in relation to the International Standard.
Dextrose 0.050
NOTE — Any of the following methods may be followed.
Maintain the bath at a temperature of 32º or at some other
Method A. By depression of the blood pressure in chicken — suitable temperature at which spontaneous contractions of
Anaesthetise a young healthy adult cockerel weighing 1.2 to the uterus are abolished and the preparation maintains its
2.3 kg with an anaesthetic that will maintain a prolonged and sensitivity. Oxygenate the solution with a mixture of 95 per
constant high blood pressure. Expose the gluteus primus cent of oxygen and 5 per cent of carbon dioxide and record
muscle in one thigh and cut and retract it to reveal the popliteal the contractions of the muscle using a suitable instrument
artery and crural vein. Cannulate the popliteal artery and record giving a linear response (for example an isotonic lever with a
the blood pressure on a suitable recorder calibrated for use load not exceeding 2 g). Record the contractions produced by
over a linear range. Cannulate the crural or brachial vein. the addition to the bath of two doses of the Standard
Immediately before use prepare a solution of the Standard Preparation suitably diluted with the above solution. The doses
Preparation in saline solution so that the volume to be injected should be such as to produce clearly discriminated,
is between 0.1 ml and 0.5 ml. Record the blood pressure submaximal contractions; the required doses normally lie
responses to the injection into the cannulated vein of two between 10 and 50 micro Units per ml of bath liquid. When
doses of this solution; the doses should be such as to produce maximal contraction has been reached, replace the bath liquid
clearly discriminated, precipitous, submaximal decreases in by a fresh solution. The doses should be added at regular
blood pressure; the required doses normally lie between 20 intervals of 3 to 5 minutes depending upon the rate of recovery
and 100 milliUnits. The interval between injections should be of the muscle. Dilute the preparation being examined so as to
constant and lie between 3 and 10 minutes depending on the obtain responses on the addition of two doses similar to those
rate at which the blood pressure returns to normal. Immediately obtained with the Standard Preparation. The ratio between
before use dilute the preparation being examined with saline the two doses of the preparation under examination should be
solution so as to obtain responses similar to those obtained the same as that between the two doses of the Standard
with the Standard Preparation. The ratio between the two doses Preparation and this ratio should be kept constant throughout
of the preparation under examination should be the same as the assay.
that between the two doses of the Standard Preparation and The two doses of Standard Preparation and the two doses of
this ratio should be kept constant throughout the assay. the preparation under examination should be given according
The two doses of the Standard Preparation and the two doses to a randomised block or a Latin square design and at least six
of the preparation under examination should be given responses to each should be recorded.
according to a randomised block or a Latin square design and Measure all the responses and calculate the result of the assay
at least six responses to each should be recorded. by standard statistical methods.
If the animal rapidly becomes insensitive to the repeated Method C. By measurement of milk-ejection pressure in a
injections of the solutions another animal must be used. lactating rat — Select a lactating rat, in the third to twenty-
Measure all the responses and calculate the result of the assay first day after parturition and weighing about 300 g, separate
by standard statistical methods. it from the litter and 30 to 60 minutes later anaesthetise (for
Method B. By contraction of the rat uterus — Inject 100 µg of example, by the intraperitoneal injection of a solution of
oestradiol benzoate intramuscularly into a female rat weighing Pentobarbitone Sodium). Tie the rat to an operating table,
120 to 200 g 18 to 24 hours before the assay. Immediately maintained at 37º, by its hind legs leaving the front legs free.
before the assay confirm by vaginal smear that the rat is in Cannulate the trachea with a short polyethylene tube of

885
OXYTOCIN INJACTION IP 2007

internal diameter about 2.5 mm in such a manner so as to Oxytocin Injection containing Oxytocin of natural origin
ensure a free airway; apply artificial respiration only if obtained by extraction and purification complies with the
necessary. Cannulate an external jugular or femoral vein with following additional requirement.
a polyethylene tube of internal diameter about 0.4 mm which Vasopressor activity. Not more than 0.5 Unit per 20 Units of
is filled with saline solution and closed with a pin. oxytocic activity, when assayed by the biological assay for
Shave the skin surrounding the inguinal and abdominal teats vasopressor activity described below.
and excise the tip of one teat, preferably the lower inguinal The vasopressor activity is estimated by comparing the activity
teat. Insert a polyethylene tube of internal diameter about of the preparation under examination with that of the Standard
0.3 mm and external diameter about 0.6 mm, to a depth sufficient Preparation of arginine vasopressin under the conditions of a
to obtain appropriate measurement of pressure (3 to 10 mm suitable method of assay.
depth), into the primary teat duct which opens onto the cut
surface and tie firmly in place with a ligature. Connect this Standard Preparation
cannula with a suitable strain gauge transducer (such as that
used for recording arterial blood pressure in the rat) and fill The Standard Preparation is the Ist International Standard for
the whole system with a 3.8 per cent w/v solution of sodium Arginine vasopressin, established in 1978, consisting of
citrate or saline solution containing 50 Units of heparin freeze-dried synthetic arginine vasopressin peptide acetate
sodium per ml to prevent clotting of milk. After cannulation, with human albumin and citric acid (supplied in ampoules
inject a small volume (0.05 to 0.2 ml) of this solution into the containing 8.20 Units), or any other suitable preparation the
teat duct through the transducer to clear the milk from the tip potency of which has been determined in relation to that of
of the cannula. (This procedure may be repeated during the the International Standard.
assay should obstruction arise from milk ejected into the Inject slowly into the tail vein of a male albino rat weighing
cannula). Clamp the strain gauge so that a slight tension is about 300 g a solution of a suitable a-adrenoceptor blocking
applied to the teat and its natural alignment is preserved and agent, for example 10 ml per kg of body weight of a solution
connect the gauge to a potentiometric recorder adjusted to prepared by dissolving 5 mg of phenoxybenzamine
give full-scale deflection for an increase in milk-ejection hydrochloride in 0.1 ml of ethanol (95 per cent), adding
pressure of about 5.3 kPa. Inject all solutions through the 0.05 ml of 1 M hydrochloric acid and diluting to 5 ml with
venous cannula using a 1-ml syringe graduated in 0.01 ml and saline solution. After 18 hours, anaesthetise the rat with an
wash them in with 0.2 ml of saline solution. anaesthetic that will maintain a prolonged and uniform blood
Prepare a solution of the Standard Preparation and a solution pressure. After 45 to 60 minutes, tie the rat on its back to the
of the preparation under examination in saline solution so operating table by its hind legs. Cannulate the trachea with a
that the volume to be injected is between 0.1 ml and 0.4 ml. short polyethylene tube of external diameter about 2.5 mm
Choose two doses of the Standard Preparation such that the and dissect a carotid artery ready for cannulation. Then
increase in milk-ejection pressure is about 1.35 kPa for the cannulate the femoral vein close to the inguinal ligament.
lower dose and about 2.7 kPa for the higher dose. As an initial Retract the abdominal muscles to expose the inguinal ligament.
approximation, a lower dose of between 0.1 and 0.4 milliUnit Retract the superficial pudendal vein to one side and dissect
and an upper dose of 1.5 to 2 times this amount may be tried. the femoral vein towards the inguinal ligament from the
Choose two doses of the preparation under examination with corresponding artery. When dissecting, a deep branch
the same inter-dose ratio, matching the effects of the doses of reaching the femoral vein must be found and tied off to prevent
the Standard Preparation as closely as possible. Inject the bleeding during cannulation. Tie a short polyethylene cannula
four doses (two doses of the Standard Preparation and two of external diameter about 1 mm into the femoral vein by two
doses of the preparation under examination at intervals of 3 to ligatures and join by a short piece of flexible tubing to a 1-ml
5 minutes. The two doses of Standard Preparation and the burette with an attached thistle funnel containing saline
two doses of the preparation under examination should be solution at about 37º. Firmly fix a wet absorbent cotton swab
given according to a randomised block or a Latin square design to the thigh so as to cover the incision and cannula. At this
and at least four responses to each should be recorded. stage inject through the venous cannula 200 Units of heparin,
dissolved in saline solution, per 100 g of body weight. Then
Measure all the responses and calculate the result of the assay tie in a carotid cannula of external diameter about 1 mm and
by standard statistical methods. connect by a column of saline solution containing heparin
The estimated potency is not less than 90 per cent and not with a suitable pressure measuring device such as a mercury
more than 111 per cent of the stated potency. The fiducial manometer of internal diameter about 2 to 3 mm.
limits of error are not less than 80 per cent and not more than The central and peripheral nervous system including both
125 per cent of the stated potency. vagus and associated sympathetic nerves is left intact. No

886
IP 2007 OXYTOCIN NASAL SOLUTION

artificial respiration is necessary. Taking care that no air is synthetic oxytocin peptide with human albumin and citric acid
injected, inject all solutions through the venous cannula by (supplied in ampoules containing 12.5 Units), or any other
means of a 1-ml syringe graduated in 0.01 ml and wash in with suitable preparation the potency of which has been determined
0.2 ml of saline solution from the burette. in relation to the International Standard.
Dilute the extract of the Standard Preparation and the NOTE — Any of the following methods may be followed.
preparation under examination with saline solution so that
Method A. By depression of the blood pressure in chicken —
the volume to be injected is between 0.1 ml and 0.5 ml.
Anaesthetise a young healthy adult cockerel weighing 1.2 to
Choose two doses of the Standard Preparation such that the 2.3 kg with an anaesthetic that will maintain a prolonged and
elevation of the blood pressure is about 4 kPa for the lower constant high blood pressure. Expose the gluteus primus
dose and about 7 kPa but always submaximal for the higher muscle in one thigh and cut and retract it to reveal the popliteal
dose, the ratio of low to high dose being determined by the artery and crural vein. Cannulate the popliteal artery and record
response and usually being 3 to 5. As an initial approximation the blood pressure on a suitable recorder calibrated for use
doses of 3 and 5 milliUnits may be tried. Choose two doses of over a linear range. Cannulate the crural or brachial vein.
the preparation under examination with the same inter-dose
Immediately before use prepare a solution of the Standard
ratio, matching the effects of the dose of the Standard
Preparation in saline solution so that the volume to be injected
Preparation as closely as possible. Inject doses at intervals of
is between 0.1 ml and 0.5 ml. Record the blood pressure
10 to 15 minutes.
responses to the injection into the cannulated vein of two
The two doses of the Standard Preparation and the two doses doses of this solution; the doses should be such as to produce
of the preparation under examination should be given in a clearly discriminated, precipitous, submaximal decreases in
randomised block or a Latin square design and four to five blood pressure; the required doses normally lie between 20
responses to each should be recorded. and 100 milliUnits. The interval between injections should be
Measure all the responses and calculate the result of the assay constant and lie between 3 and 10 minutes depending on the
by standard statistical methods. rate at which the blood pressure returns to normal. Immediately
before use dilute the preparation being examined with saline
Storage. Store at temperature not exceeding 30º. Do not freeze. solution so as to obtain responses similar to those obtained
Labelling. The label states (1) the number of Units of oxytocin with the Standard Preparation. The ratio between the two doses
activity per ml; (2) either the animal spieces from which it is of the preparation under examination should be the same as
obtained or whether it is synthetic, as appropriate. that between the two doses of the Standard Preparation and
this ratio should be kept constant throughout the assay.
The two doses of the Standard Preparation and the two doses
Oxytocin Nasal Solution of the preparation under examination should be given
according to a randomised block or a Latin square design and
Oxytocin Nasal Solution is a solution of Oxytocin in a suitable
at least six responses to each should be recorded.
solvent containing an appropriate antimicrobial preservative.
If the animal rapidly becomes insensitive to the repeated
Oxytocin Nasal Solution contains not less than 85.0 per cent
injections of the solutions another animal must be used.
and not more than 120.0 per cent of the stated number of Units
Measure all the responses and calculate the result of the assay
of oxytocic activity.
by standard statistical methods.
Description. A clear, colourless solution.
Method B. By contraction of the rat uterus — Inject 100 µg of
Tests oestradiol benzoate intramuscularly into a female rat weighing
120 to 200 g 18 to 24 hours before the assay. Immediately
pH (2.4.24). 3.5 to 4.5. before the assay confirm by vaginal smear that the rat is in
Other Tests. Complies with the tests stated under Nasal oestrus or preoestrus. Kill the rat and suspend one horn of
Preparations. the uterus in a bath containing a solution of the following
Assay.The potency of oxytocin is determined by comparing composition.
its activity with that of the Standard Preparation of oxytocin Composition (per cent w/v)
under the conditions of a suitable method of assay. Sodium chloride 0.662
Standard Preparation Potassium chloride 0.045
The Standard Preparation is the 4th International Standard for Calcium chloride 0.007
Oxytocin, established in 1978, consisting of freeze-dried Sodium bicarbonate 0.256

887
OXYTOCIN NASAL SOLUTION IP 2007

Disodium hydrogen phosphate 0.029 to obtain appropriate measurement of pressure (3 to 10 mm


Sodium dihydrogen phosphate 0.003 depth), into the primary teat duct which opens onto the cut
surface and tie firmly in place with a ligature. Connect this
Magnesium chloride 0.010
cannula with a suitable strain gauge transducer (such as that
Dextrose 0.050 used for recording arterial blood pressure in the rat) and fill
Maintain the bath at a temperature of 32º or at some other the whole system with a 3.8 per cent w/v solution of sodium
suitable temperature at which spontaneous contractions of citrate or saline solution containing 50 Units of heparin
the uterus are abolished and the preparation maintains its sodium per ml to prevent clotting of milk. After cannulation,
sensitivity. Oxygenate the solution with a mixture of 95 per inject a small volume (0.05 to 0.2 ml) of this solution into the
cent of oxygen and 5 per cent of carbon dioxide and record teat duct through the transducer to clear the milk from the tip
the contractions of the muscle using a suitable instrument of the cannula. (This procedure may be repeated during the
giving a linear response (for example an isotonic lever with a assay should obstruction arise from milk ejected into the
load not exceeding 2 g). Record the contractions produced by cannula). Clamp the strain gauge so that a slight tension is
the addition to the bath of two doses of the Standard applied to the teat and its natural alignment is preserved and
Preparation suitably diluted with the above solution. The doses connect the gauge to a potentiometric recorder adjusted to
should be such as to produce clearly discriminated, give full-scale deflection for an increase in milk-ejection
submaximal contractions; the required doses normally lie pressure of about 5.3 kPa. Inject all solutions through the
between 10 and 50 micro Units per ml of bath liquid. When venous cannula using a 1-ml syringe graduated in 0.01 ml and
maximal contraction has been reached, replace the bath liquid wash them in with 0.2 ml of saline solution.
by a fresh solution. The doses should be added at regular Prepare a solution of the Standard Preparation and a solution
intervals of 3 to 5 minutes depending upon the rate of recovery of the preparation under examination in saline solution so
of the muscle. Dilute the preparation being examined so as to that the volume to be injected is between 0.1 ml and 0.4 ml.
obtain responses on the addition of two doses similar to those Choose two doses of the Standard Preparation such that the
obtained with the Standard Preparation. The ratio between increase in milk-ejection pressure is about 1.35 kPa for the
the two doses of the preparation under examination should be lower dose and about 2.7 kPa for the higher dose. As an initial
the same as that between the two doses of the Standard approximation, a lower dose of between 0.1 and 0.4 milliUnit
Preparation and this ratio should be kept constant throughout and an upper dose of 1.5 to 2 times this amount may be tried.
the assay. Choose two doses of the preparation under examination with
The two doses of Standard Preparation and the two doses of the same inter-dose ratio, matching the effects of the doses of
the preparation under examination should be given according the Standard Preparation as closely as possible. Inject the
to a randomised block or a Latin square design and at least six four doses (two doses of the Standard Preparation and two
responses to each should be recorded. doses of the preparation under examination at intervals of 3 to
5 minutes. The two doses of Standard Preparation and the
Measure all the responses and calculate the result of the assay
two doses of the preparation under examination should be
by standard statistical methods.
given according to a randomised block or a Latin square design
Method C. By measurement of milk-ejection pressure in a and at least four responses to each should be recorded.
lactating rat — Select a lactating rat, in the third to twenty- Measure all the responses and calculate the result of the assay
first day after parturition and weighing about 300 g, separate by standard statistical methods.
it from the litter and 30 to 60 minutes later anaesthetise (for
example, by the intraperitoneal injection of a solution of The estimated potency is not less than 90 per cent and not
Pentobarbitone Sodium). Tie the rat to an operating table, more than 111 per cent of the stated potency. The fiducial
maintained at 37º, by its hind legs leaving the front legs free. limits of error are not less than 80 per cent and not more than
Cannulate the trachea with a short polyethylene tube of 125 per cent of the stated potency.
internal diameter about 2.5 mm in such a manner so as to Oxytocin Nasal Solution containing Oxytocin of natural
ensure a free airway; apply artificial respiration only if origin obtained by extraction and purification complies with
necessary. Cannulate an external jugular or femoral vein with the following additional requirement.
a polyethylene tube of internal diameter about 0.4 mm which
is filled with saline solution and closed with a pin. Vasopressor activity. Not more than 0.5 Unit per 20 Units of
oxytocic activity, when assayed by the biological assay for
Shave the skin surrounding the inguinal and abdominal teats
vasopressor activity described below.
and excise the tip of one teat, preferably the lower inguinal
teat. Insert a polyethylene tube of internal diameter about The vasopressor activity is estimated by comparing the activity
0.3 mm and external diameter about 0.6 mm, to a depth sufficient of the preparation under examination with that of the Standard

888
IP 2007 OXYTOCIN NASAL SOLUTION

Preparation of arginine vasopressin under the conditions of a tie in a carotid cannula of external diameter about 1 mm and
suitable method of assay. connect by a column of saline solution containing heparin
with a suitable pressure measuring device such as a mercury
Standard Preparation manometer of internal diameter about 2 to 3 mm.
The Standard Preparation is the Ist International Standard for The central and peripheral nervous system including both
Arginine vasopressin, established in 1978, consisting of vagus and associated sympathetic nerves is left intact. No
freeze-dried synthetic arginine vasopressin peptide acetate artificial respiration is necessary. Taking care that no air is
with human albumin and citric acid (supplied in ampoules injected, inject all solutions through the venous cannula by
containing 8.20 Units), or any other suitable preparation the means of a 1-ml syringe graduated in 0.01 ml and wash in with
potency of which has been determined in relation to that of 0.2 ml of saline solution from the burette.
the International Standard.
Dilute the extract of the Standard Preparation and the
Inject slowly into the tail vein of a male albino rat weighing preparation under examination with saline solution so that
about 300 g a solution of a suitable a-adrenoceptor blocking the volume to be injected is between 0.1 ml and 0.5 ml.
agent, for example 10 ml per kg of body weight of a solution
Choose two doses of the Standard Preparation such that the
prepared by dissolving 5 mg of phenoxybenzamine
elevation of the blood pressure is about 4 kPa for the lower
hydrochloride in 0.1 ml of ethanol (95 per cent), adding
dose and about 7 kPa but always submaximal for the higher
0.05 ml of 1 M hydrochloric acid and diluting to 5 ml with
dose, the ratio of low to high dose being determined by the
saline solution. After 18 hours, anaesthetise the rat with an
response and usually being 3 to 5. As an initial approximation
anaesthetic that will maintain a prolonged and uniform blood
doses of 3 and 5 milliUnits may be tried. Choose two doses of
pressure. After 45 to 60 minutes, tie the rat on its back to the
the preparation under examination with the same inter-dose
operating table by its hind legs. Cannulate the trachea with a
ratio, matching the effects of the dose of the Standard
short polyethylene tube of external diameter about 2.5 mm
Preparation as closely as possible. Inject doses at intervals of
and dissect a carotid artery ready for cannulation. Then
10 to 15 minutes.
cannulate the femoral vein close to the inguinal ligament.
Retract the abdominal muscles to expose the inguinal ligament. The two doses of the Standard Preparation and the two doses
Retract the superficial pudendal vein to one side and dissect of the preparation under examination should be given in a
the femoral vein towards the inguinal ligament from the randomised block or a Latin square design and four to five
corresponding artery. When dissecting, a deep branch responses to each should be recorded.
reaching the femoral vein must be found and tied off to prevent Measure all the responses and calculate the result of the assay
bleeding during cannulation. Tie a short polyethylene cannula by standard statistical methods.
of external diameter about 1 mm into the femoral vein by two
ligatures and join by a short piece of flexible tubing to a 1-ml Storage. Store at a temperature not exceeding 30º.
burette with an attached thistle funnel containing saline Labelling. The label states (1) the number of Units of oxytocic
solution at about 37º. Firmly fix a wet absorbent cotton swab activity per ml; (2) either the animal species from which it is
to the thigh so as to cover the incision and cannula. At this obtained or whether it is synthetic, as appropriate; (3) that the
stage inject through the venous cannula 200 Units of heparin, preparation is intended for intranasal administration only.
dissolved in saline solution, per 100 g of body weight. Then

889
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

P
Paclitaxel ....
Paclitaxel Injection ....
Pancreatin ....
D-Panthenol ....
Paracetamol ....
Paracetamol Syrup ....
Paracetamol Tablets ....
Hard Paraffin ....
Liquid Paraffin ....
Light Liquid Paraffin ....
Liquid Paraffin Emulsion ....
White Soft Paraffin ....
Yellow Soft Paraffin ....
Paraffin Ointment ....
Paraldehyde ....
Penicillamine ....
Penicillamine Tablets ....
Diluted Pentaerythritol Tetranitrate ....
Pentaerythritol Tetranitrate Tablets ....
Pentamidine Isethionate ....
Pentamidine Injection ....
Pentazocine ....
Pentazocine Hydrochloride ....
Pentazocine Tablets ....
Pentazocine Lactate ....
Pentazocine Injection ....
Pentobarbitone Sodium ....
Pentobarbitone Tablets ....
Pepsin ....
Peritoneal Dialysis Solutions ....

891
MONOGRAPHS INDIAN PHARMACOPOEIA 2007 2007

Pethidine Hydrochloride
Pethidine Injection
Pethidine Tablets
Phenindamine Tartrate
Phenindamine Tablets
Phenindione
Phenindione Tablets
Pheniramine Maleate
Pheniramine Injection
Pheniramine Tablets
Phenobarbitone
Phenobarbitone Tablets
Phenobarbitone Sodium
Phenobarbitone Injection
Phenobarbitone Sodium Tablets
Phenol
Phenolphthalein
Phenoxymethylpenicillin Potassium
Phenoxymethylpenicillin Potassium Tablets
Phentolamine Mesylate
Phentolamine Injection
Phenylbutazone
Phenylbutazone Tablets
Phenylephrine Hydrochloride
Phenylephrine Injection
Phenylmercuric Acetate
Phenylmercuric Nitrate
Phenytoin Sodium
Phenytoin Injection
Phenytoin Tablets
Pholcodine
Pholcodine Linctus
Phosphoric Acid

892
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

Physostigmine Salicylate ....


Physostigmine Injection ....
Pilocarpine Nitrate ....
Pindolol ....
Pindolol Tablets ....
Piperazine Adipate ....
Piperazine Adipate Tablets ....
Piperazine Citrate ....
Piperazine Citrate Syrup ....
Piperazine Hydrate ....
Piperazine Phosphate ....
Piperazine Phosphate Tablets ....
Piroxicam ....
Piroxicam Capsules ....
Plaster Of Paris ....
Polyethylene Glycol 1500 ....
Polyethylene Glycol 4000 ....
Polyethylene Glycol 6000 ....
Polysorbate 20 ....
Polysorbate 80 ....
Potassium Chloride ....
Potassium Chloride And Dextrose Injection ....
Potassium Chloride, Sodium Chloride And Dextrose Injection ....
Potassium Citrate ....
Potassium Clavulanate ....
Potassium Clavulanate Diluted ....
Potassium Iodide ....
Potassium Permanganate ....
Povidone ....
Povidone-Iodine ....
Povidone-Iodine Solution ....
Pralidoxime Chloride ....

893
MONOGRAPHS INDIAN PHARMACOPOEIA 2007 2007

Pralidoxime Chloride Injection ....


Prazosin Hydrochloride ....
Prazosin Tablets ....
Prednisolone ....
Prednisolone Tablets ....
Prednisone ....
Prednisone Tablets ....
Prednisolone Sodium Phosphate ....
Prednisolone Sodium Phosphate Injection ....
Primaquine Phosphate ....
Primaquine Tablets ....
Probenecid ....
Probenecid Tablets ....
Procainamide Hydrochloride ....
Procainamide Injection ....
Procainamide Tablets ....
Procaine Hydrochloride ....
Procaine And Adrenaline Injection ....
Procaine Penicillin ....
Fortified Procaine Penicillin Injection ....
Prochlorperazine Maleate ....
Prochlorperazine Tablets ....
Prochlorperazine Mesylate ....
Prochlorperazine Injection ....
Procyclidine Hydrochloride ....
Procyclidine Tablets ....
Proguanil Hydrochloride ....
Proguanil Tablets ....
Promethazine Hydrochlorid ....
Promethazine Injection ....
Promethazine Syrup ....
Promethazine Tablets ....
Promethazine Theoclate ....

894
INDIAN PHARMACOPOEIA 2007 MONOGRAPHS

Promethazine Theoclate Tablets ....


Propantheline Bromide ....
Propantheline Tablets ....
Propranolol Hydrochloride ....
Propranolol Injection ....
Propranolol Tablets ....
Propyl Gallate ....
Propylene Glycol ....
Propylparaben ....
Propylthiouracil ....
Propylthiouracil Tablets ....
Propyphenazone ....
Protamine Sulphate ....
Protamine Sulphate Injection ....
Prothionamide ....
Prothionamide Tablets ....
Pseudoephedrine Hydrochloride ....
Pseudoephedrine Syrup ....
Pseudoephedrine Tablets ....
Psoralen ....
Pyrazinamide ....
Pyrazinamide Tablets ....
Pyridoxine Hydrochloride ....
Pyridoxine Tablets ....
Pyrimethamine ....
Pyrimethamine And Sulphadoxine Tablets ....

895
IP 2007 PACLITAXEL

Paclitaxel – mobile phase: A. a mixture of 60 volumes of water and


40 volumes of acetonitrile,
Taxol B. acetonitrile
– flow rate. 1.2 ml per minute,
O – a linear gradient programme using the conditions given
O below,
O
H3C – spectrophotometer set at 227nm,
CH3 – a 10 µl loop injector.
H3C CH3 OH
O NH O CH3 Time Mobile phase A Mobile phase B
(in min) (per cent v/v) (per cent v/v)
O O
OH H 0 100 0
OH O O
CH3 20 100 0
O O 60 10 90
62 100 0
C47H51NO14 Mol. Wt. 853.9 70 100 0
A taxane derivative first isolated from the bark of the Pacific Inject reference solution (b). The test is not valid unless the
yew tree, Taxus brevifolia. resolution between the peak due to paclitaxel and 10-dactyl -
7-epipactitaxyl is not less than 1.2 relative retention time for
Paclitaxel contains not less than 97.0 per cent and not more
particles and 10-dactyl -7-epipactitaxyl is about 0.94.
than 102.0 per cent of C47H51NO14, calculated on the anhydrous
basis. Inject the test solution and reference solution (a). In the
chromatogram obtained with the test solution, the area of any
Description. A white or almost white powder. secondary peak is not more than 0.5 times the area of the peak
CAUTION — Paclitaxel is potentially cytotoxic. Great care in the chromatogram obtained with the reference solution (a)
should be taken in handling the powder and preparing (0.5 per cent) and the sum of areas of all the secondary peaks
solutions. is not more than twice the area of the peak in the chromatogram
obtained with the reference solution (a) (2.0 per cent).
Identification
Heavy metals. (2.3.13). 1 g complies with the limit test for heavy
A. Determine by infrared absorption spectrophotometry (2.4.6). metals, Method A ( 20ppm ).
Compare the spectrum with that obtained with paciltaxel RS.
Loss on ignition (2.4.20). Not more than 0.2 per cent.
B. In the Assay, the principal peak in the chromatogram Water (2.3.43). Not more than 4.0 per cent, determined on 0.1
obtained with the test solution corresponds to the peak in the g by coloumetry method.
chromatogram obtained with the reference solution.
Bacterial endotoxins (2.2.3). Not more than 0.4 Endotoxin Unit
Tests per mg of paclitaxel.
Specific optical rotation (2.4.22). – 49.0º to – 55.0º, determined Microbial contamination (2.2.9). The total viable aerobic count
in 1.0 per cent w/v solution in methanol. does not exceed 100 cfu per g. It meets the requirements of the
tests for the absence of Staphylococcus aureus, Pseudomonas
Related substances. Determine by liquid chromatography
aeruginosa, Salmonella species, and Escherichia coli.
(2.4.14).
Assay. Determine by liquid chromatography (2.4.14).
Test solution. Dissolve 10 mg of substance under examination
in 10 ml of acetonitrile. Solvent mixture. Dissolve 200 µl of glacial acetic acid in
1000 ml of methanol.
Reference solution (a). A 0.001 per cent w/v solution of
paclitaxel RS in acetonitrile. Test solution. Dissolve 0.1 g of the substance under
examination in 100.0 ml in solvent mixture.
Reference solution (b). A sloution containing 0.008 per cent
w/v of 10 deacetyl -7-epipoelitoral and 0.1 per cent w/v of Reference solution. A 0.1 per cent w/v solution of paclitaxil
paclitaxel RS in acetonitrile. RS in solvent mixture.
Chromatographic system Chromatographic system
– a stainless steel column 15 cm x 4.6 mm, packed with – a stainless steel column 25 cm x 4.6 mm packed with
octadecylsilane bonded to porous silica (3µm). pentafluoro phenyl groups chemically bonded to
– column temperature 35°, porous silica (5 µm),

897
PACLITAXEL INJECTION IP 2007

– column temperature 35°, System suitability solution- A sloution containing 0.006 per
– mobile phase: a mixture of 11 volumes of water and 9 cent w/v of 10 deacetyl -7-epipoelitoral and 0.12 per cent w/v
volumes of acetonitrile, filter. of paclitaxel in acetonitrile.
– flow rate. 1.5 ml per minute, Chromatographic system
– spectrophotometer set at 227nm, – a stainless steel column 15 cm x 4.6 mm, packed with
– a 10 µl loop injector. octadecylsilane bonded to porous silica (3 µm).
Inject the reference solution. The test is not valid unless the – column temperature 35°,
tailing factor is not more than 1.5. The relative standard – mobile phase: A. a mixture of 60 volumes of water and
deviation for replicate injections is not more than 2.0 per cent. 40 volumes of acetonitrile,
B. acetonitrile
Inject the test solution and reference solution.
– flow rate. 1.2 ml per minute,
Calculate the content of C47H51NO14. – a linear gradient programme using the conditions given
Storage. Store protected from light, at a temperature not below,
exceeding 25º. – spectrophotometer set at 227nm,
– a 10 µl loop injector.
Time Mobile phase A Mobile phase B
(in min) (per cent v/v) (per cent v/v)
Paclitaxel Injection 0 100 0
26 100 0
Paclitaxel Injection is a sterile solution of Paclitaxel suitable
for dilution,for intravenous use. 66 17 83
67 100 0
Paclitaxel Injection contains not less than 90.0 per cent and
not more than 105.0 per cent of of the related amount of Inject reference solution (b). The test is not valid unless the
paclitaxel, C47H51NO14. column efficiency is not less than 2000 theoretical plates and
the tailing factor is not more than 2.0.
Description. A clear colourless to slight yellow viscous
solution. Inject the test solution and reference solution (a). In the
chromatogram obtained with the test solution, the area of any
Identification secondary peak is not more than 0.8 times the area of the peak
in the chromatogram obtained with the reference solution (a)
A. In the Related substances, the major peak in the (0.8 per cent) and the sum of areas of all the secondary peaks
chromatogram obtained with test solution corresponds to the is not more than twice the area of the peak in the chromatogram
peak in the chromatogram obtained with reference solution. obtained with the reference solution (a) (2.0 per cent).
B. In the Assay, the principal peak in the chromatogram Other tests. Complies with the tests stated under Parenteral
obtained with test solution corresponds to the peak in the Preparations (Injections).
chromatogram obtained with reference solution.
Bacterial endotoxins (2.2.3). Not more than 0.67 Endotoxin
Tests Unit per mg of paclitaxel.
Sterility (2.2.11). Complies with the test for sterility.
pH (2.4.24). 3.0 to 7.0, determined in a solution 10 per cent v/v
solution in water. Assay. Determine by liquid chromatography (2.4.14).

Light absorption. Absorbance of the injection at about 425 Solvent mixture. Dissolve 200 µl of glacial acetic acid in
nm, not more than 0.01. 1000 ml of methanol.
Test solution. Accurately measure the volume of injection
Related substances. Determine by liquid chromatography
containing 6 mg of Paclitaxel and dissolve in 10 ml of solvent
(2.4.14).
mixture.
Test solution. Accurately measure the volume of injection
Reference solution. A 0.06 per cent w/v solution of paclitaxel
containing 12 mg of Paclitaxel, dilute to 10 ml with acetonitrile.
RS in solvent mixture.
Reference solution (a). A 0.12 per cent w/v solution of
Chromatographic system
paclitaxel RS in aceotnitrile.
– a stainless steel column 30 cm x 3.9 mm, packed with
Reference solution (b). Dilute 1 ml of reference solution (a) to penta fluro phenyl group chemically bonded to porous
100 ml with acetonitrile. silica (5 µm),

898
IP 2007 PANCREATIN

– column temperature 35°, Tests


– mobile phase: a mixture of 11 volumes of water and 9
volumes of acetonitrile, Fat. Not more than 5.0 per cent, determined by the following
– flow rate. 1.5 ml per minute, method. Extract 1 g with light petroleum (40º to 60º) for
– spectrophotometer set at 227nm, 3 hours in an apparatus for the continuous extraction of drugs
– a 10 µl loop injector. (2.1.8), evaporate the extract and dry the residue at 105º for
2 hours.
Inject the reference solution. The test is not valid unless the
tailing factor is not more than 1.5. The relative standard Microbial contamination (2.2.9). 1 g is free from Escherichia
deviation for replicate injections is not more than 2.0 per cent. coli; 10 g is free from salmonellae.

Inject the test solution and reference solution. Loss on drying (2.4.19). Not more than 5.0 per cent, determined
on 0.5 g by dying in an oven at 60º at a pressure not exceeding
Calculate the content of C47H51NO14. 0.7 kPa for 4 hours.
Storage. Store protected from light, at a temperature not Assay. For protease activity — Weigh accurately 4.0 g of
exceeding 25º. purified casein and dissolve in about 90 ml of water containing
3 ml of 1 M sodium hydroxide, adjust the pH of the solution to
8.7 and add sufficient water to produce 100.0 ml. Weigh
accurately about 0.5 g of the substance under examination,
Pancreatin triturate with water and add sufficient water to produce
300.0 ml to give the test solution. Dilute 15.0 ml of the casein
Pancreatin is a preparation of mammalian pancreas containing solution with 30 ml of water, warm to 55º and add 10.0 ml of the
protease, lipase and amylase activity. It may contain Sodium unfiltered test solution. Heat rapidly to 55º and keep at this
Chloride. temperature for 20 minutes. Cool rapidly to room temperature.
Dilute a further portion of 15.0 ml of the casein solution with
Pancreatin contains not less than the minimum protease
30 ml of water, add 10.0 ml of the unfiltered test solution,
activity, amylase activity and lipase activity determined under
previously boiled and cooled, heat rapidly to 55º and keep at
the conditions of the Assay.
this temperature for 20 minutes. Cool to room temperature. To
Description. A white or buff-coloured, amorphous powder; each solution add 0.75 ml of phenolphthalein solution and 10
odour, meaty and not unpleasant. ml of formaldehyde solution. Titrate each solution with 0.1 M
sodium hydroxide until the colour of the solution matches
Identification that produced by mixing 10 ml of buffer solution pH 8.7 and
0.15 ml of phenolphthalein solution. The difference between
A. Triturate 0.5 g with 10 ml of water and adjust to pH 8.0 by
the two titrations is not less than 4.5 ml.
the addition of 1 M sodium hydroxide using cresol red
solution as indicator. Divide the resulting solution into two For lipase activity — To 95 ml of water add 6.5 ml of triacetin
equal portions. Boil one portion [solution (1)] and leave the and 0.2 ml of a 0.1 per cent w/v solution of bromocresol purple,
other untreated [solution (2)]. To each add a few shreds of neutralise with 0.5 M sodium hydroxide and add sufficient
congo red fibrin, warm to 39º ± 1º and maintain at this water to produce 110 ml. Place 50 ml of this solution in each of
temperature for 1 hour. Solution (2) is stained red and solution two large tubes 3 cm ´ 20 cm A and B contained in a thermostat
(1) is colourless or not more than slightly pink. at 30º. Insert in each tube a rubber stopper having two holes,
one for the tip of a burette and the other for a short glass tube
B. Triturate 0.25 g with 10 ml of water and adjust to pH 8.0 by
through which passes a thread operating a glass stirring coil.
the addition of 1 M sodium hydroxide using cresol red
Stir the contents of the tube until they attain the temperature
solution as indicator. Divide the resulting solution into two
of the thermostat. Prepare a solution of 0.1 g of the substance
equal portions. Boil one portion [solution (1)] and leave the
under examination in 10.0 ml of water. To tube A add 1.0 ml of
other untreated [solution (2)]. Dissolve 0.1 g of soluble starch
the solution, to tube B add 1.0 ml of the solution previously
in 100 ml of boiling water, boil for 2 minutes, cool and dilute to
boiled. Adjust and maintain the pH of the solutions in the two
150 ml with water. Add solution (1) to half the starch mucilage
tubes to 6.2 to 6.4 by the addition of 0.05 M sodium hydroxide
and solution (2) to the remainder and maintain the mixtures at
dropwise, stirring frequently. After 30 minutes, the difference
39º ± 1º for 5 minutes. To 1 ml of each mixture add 10 ml of
between the volumes of 0.05 M sodium hydroxide added to
iodinated potassium iodide solution. The liquid containing
the two tubes is not less than 1.0 ml.
solution (2) retains the colour of the solution of iodine and
the liquid containing solution (1) acquires an intense blue For amylase activity — Not less than 100 Units per g. Dissolve
colour. 0.1 g or a quantity containing 10 Units, accurately weighed, in

899
D-PANTHENOL IP 2007

sufficient buffer solution pH 6.8 to produce 1000.0 ml. Filter if Refractive index (2.4.27). 1.490 to 1.498, determined at 20º.
necessary (1 ml of the test solution should be capable of Heavy metals (2.3.13). 1.0 g dissolved in 25 ml of water complies
digesting about 10 mg of dry soluble maize or corn starch). with the limit test for heavy metals, Method A (20 ppm).
Into each of six stoppered test-tubes add 5.0 ml of starch
substrate without touching the sides of the test-tube. Place Assay. Weigh accurately about 0.5 g and carry out Method A
the test-tubes in a water-bath maintained at 40º ± 0.1º. When for the determination of nitrogen (2.3.30).
the temperature of the solution in the tubes has reached 40º, 1 ml of 0.05 M sulphuric acid is equivalent to 0.001401 g of N.
add 0.35 ml, 0.4 ml, 0.45 ml, 0.5 ml, 0.55 ml and 0.6 ml of the test
solution to each of the test-tubes marked 1 to 6 respectively Storage. Store protected from moisture.
and record the time of addition. Mix thoroughly and replace
the tubes in the water-bath. After exactly 60 minutes remove
the tubes and cool rapidly in cold water. Add to each tube Paracetamol
0.05 ml of 0.02 M iodine and mix well. Note the tube containing
the lowest volume of test solution, which does not show a Acetaminophen
bluish or violet tinge (if there is doubt, warm the solution
slightly, when the colour distinction is prominent). From this O
volume calculate the number of grams of dry soluble maize or
HN CH3
corn starch digested by 1.0 g of the substance under
examination. This represents the number of Units of amylase
activity per g.
Storage. Store protected from moisture.
OH
Labelling. The label states the name of any added substance.
C8H9NO2 Mol. Wt. 151.2
Paracetamol is 4-hydroxyacetanilide.
D-Panthenol Paracetamol contains not less than 99.0 per cent and not more
Pantothenol; Dextro-pantothenyl Alcohol than 101.0 per cent of C8H9NO2, calculated on the dried basis.
Description. White crystals or a white, crystalline powder.
H OH
HO N OH Identification
H3C CH3 O H Test A may be omitted if tests B C and D are carried out. Tests
C9H19NO4 Mol. Wt. 205.3 B, C and D may be omitted if test A is carried out.
A. Determine by infrared absorption spectrophotometry (2.4.6).
D-Panthenol is (R)-2,4-dihydroxy-N-(3-hydroxypropyl)-3, 3-
Compare the spectrum with that obtained with paracetamol
dimethylbutanamide.
RS or with the reference spectrum of paracetamol.
D-Panthenol contains not less than 6.60 per cent and not more
B. Dissolve 50 mg in sufficient methanol to produce 100 ml.
than 6.95 per cent of nitrogen, N.
To 1 ml of this solution add 0.5 ml of 0.1 M hydrochloric acid
Description. A clear, colourless or slightly yellow, viscous and dilute to 100 ml with methanol. Protect the resulting
liquid; odourless. solution from bright light and immediately measure the
absorbance at the maximum at about 249 nm; absorbance at
Identification 249 nm, about 0.44 (2.4.7).
Boil 50 mg with 5 ml of 0.1 M sodium hydroxide for 1 minute, C. Boil 0.1 g in 1 ml of hydrochloric acid for 3 minutes, add
cool and add 5 ml of 1 M hydrochloric acid and 0.1 ml of ferric 10 ml of water and cool; no precipitate is produced. Add
chloride test solution; a deep yellow colour is produced. 0.05 ml of 0.0167 M potassium dichromate; a violet colour
develops which does not turn red.
Tests
D. Gives the reaction of acetyl groups (2.3.1).
pH (2.4.24). Not more than 10.5, determined in a 5.0 per cent
w/v solution in carbon dioxide-free water. Tests
Specific optical rotation (2.4.22). +28.2º to +30.2º, determined 4-Aminophenol. Dissolve 0.5 g in sufficient methanol (50 per
at 20º in a 5.0 per cent w/v solution. cent) to produce 10 ml. Add 0.2 ml of freshly prepared alkaline

900
IP 2007 PARACETAMOL SYRUP

sodium nitroprusside solution, mix and allow to stand for 1 ml of 0.1 M ceric ammonium sulphate is equivalent to
30 minutes. Any blue colour in the solution is not more intense 0.00756 g of C8H9No2.
than that in 10 ml of a solution prepared at the same time and Storage. Store protected from light and moisture.
in the same manner containing 0.5 g of 4-aminophenol-free
paracetamol and 0.5 ml of a 0.005 per cent w/v solution of
4-aminophenol in methanol (50 per cent) (50 ppm).
Related substances. Determine by thin layer chromatograpy Paracetamol Syrup
(2.4.17), coating the plate with silica gel GF254. Paracetamol Oral Solution; Acetaminophen Syrup
Mobile phase. A mixture of 65 volumes of chloroform, Paracetamol Syrup is a solution of Paracetamol in a suitable
25 volumes of acetone and 10 volumes of toluene. flavoured vehicle.
Test solution (a). Transfer 1 g of the substance under Paracetamol Syrup contains not less than 95.0 per cent and
examination, finely powdered, to a ground-glass stoppered not more than 105.0 per cent w/v solution of the stated amount
15-ml centrifuge tube, add 5 ml of peroxide-free ether, shake of paracetamol, C8H9NO2.
mechanically for 30 minutes and centrifuge at 1000 rpm for 15
minutes or until a clear supernatant liquid is obtained. Identification
Test solution (b). Dilute 1 ml of the test solution to 10 ml with Determine by thin-layer chromatography (2.4.17), coating the
ethanol (95 per cent). plate with silica gel GF254.
Reference solution (a). A 0.005 per cent w/v solution of Mobile phase. A mixture of 65 volumes of chloroform,
4-chloroacetanilide in ethanol (95 per cent). 25 volumes of acetone, 10 volumes of toluene and 0.5 volumes
of glacial acetic acid.
Reference solution (b). Dissolve 0.25 g of 4-chloroacetanilide
and 0.1 g of the substance under examination in sufficient Test solution. Dilute a volume containing 25 mg of Paracetamol
ethanol (95 per cent) to produce 100 ml. to 10 ml with methanol and filter if necessary.
Apply to the plate 200 µl of test solution (a) and 40 µl of each Reference solution. A 0.25 per cent w/v solution of
of test solution (b) and reference solutions (a) and (b). After paracetamol RS in methanol
development, dry the plate in a current of warm air and examine Apply to the plate 10 µl of each solution. After development,
in ultraviolet light at 254 nm. Any spot corresponding to dry the plate in a current of warm air, examine in ultraviolet
4-chloroacetanilide in the chromatogram obtained with test light at 254 nm. The principal spot in the chromatogram
solution (a) is not more intense than the spot in the obtained with the test solution corresponds to that in the
chromatogram obtained with reference solution (a). Any other chromatogram obtained with the reference solution.
secondary spot in the chromatogram obtained with test
solution (b) is not more intense than the spot in the Tests
chromatogram obtained with reference solution (a). The 4-Aminophenol. Determine by liquid chromatography (2.4.14).
chromatogram obtained with reference solution (b) shows two
clearly separated spots, the spot corresponding to paracetamol Test solution. Shake 5 ml of the preparation under examination
having the lower Rf value. with 15 ml of the mobile phase, dilute to 25 ml with the mobile
phase and filter if necessary.
Heavy metals (2.3.13). 2.0 g complies with the limit test for
heavy metals, Method B (10 ppm). Reference solution. A 0.0025 per cent w/v solution of
4-aminophenol in the mobile phase.
Sulphated ash (2.3.18). Not more than 0.1 per cent.
Chromatographic system
Loss on drying (2.4.19). Not more than 0.5 per cent, determined – a stainless steel column 20 cm x 4.6 mm, packed with
on 1.0 g by drying in an oven at 105º. octadecylsilane bonded to porous silica (10 µm),
– mobile phase: 0.01 M sodium butanesulphonate in a
Assay. Weigh accurately about 0.5 g, dissolve in a mixture of
mixture of 85 volumes of water, 15 volumes of methanol
10 ml of water and 50 ml of 1 M sulphuric acid. Boil under a
and 0.4 volume of formic acid,
reflux condenser for 1 hour, cool and dilute to 100.0 ml with
– flow rate. of 2 ml per minute,
water. To 20.0 ml of the solution add 40 ml of water, 40 g of
– spectrophotometer set at 272 nm,
water in the form of ice, 15 ml of 2 M hydrochloric acid and
– a 20 µl loop injector.
0.1 ml of ferroin solution and titrate with 0.1 M ceric
ammonium sulphate until a yellow colour is produced. Carry In the chromatogram obtained with the test solution the area
out a blank titration. of any peak corresponding to 4-aminophenol is not greater

901
PARACETAMOL TABLETS IP 2007

than the area of the peak in the chromatogram obtained with Test solution. Shake a quantity of the powdered tablets
the reference solution. In the chromatogram obtained with the containing 1 g of Paracetamol with 15 ml of methanol, dilute to
test solution peaks with a long retention time may occur due 100 ml with water and filter.
to preservatives in the preparations.
Reference solution. A 0.001 per cent w/v solution of
Other tests. Complies with the tests stated under Oral Liquids. 4-aminophenol in methanol (15 per cent).
Assay. Determine by liquid chromatography (2.4.14). Chromatographic system
Test solution. Mix an accurately weighed quantity of the – a stainless steel column 20 cm x 4.6 mm, packed with
preparation under examination containing 25 mg of octadecylsilane bonded to porous silica (10 µm),
Paracetamol in 100 ml of the mobile phase, dilute to 200.0 ml – mobile phase: 0.01 M sodium butanesulphonate in a
with the mobile phase and filter if necessary. mixture of 85 volumes of water, 15 volumes of methanol
and 0.4 volume of formic acid,
Reference Solution. A 0.0125 per cent w/v solution of – flow rate. 2 ml per minute,
paracetamol RS in the mobile phase. – spectrophotometer set at 272 nm,
Chromatographic system – a 20 µl loop injector.
– a stainless steel column 20 cm x 4.6 mm, packed with In the chromatogram obtained with the test solution the area
octadecylsilane bonded to porous silica (10 µm), of any peak corresponding to 4-aminophenol is not greater
– mobile phase: 0.01 M sodium butanesulphonate in a than the area of the peak in the chromatogram obtained with
mixture of 85 volumes of water, 15 the reference solution. In the chromatogram obtained with the
– volumes of methanol and 0.4 volume of formic acid, test solution peaks with a long retention times may occur due
– flow rate. of 2 ml per minute, to excipients.
– spectrophotometer set at 243 nm,
– a 20 µl loop injector. Related substances. Determine by thin layer chromatograpy
(2.4.17), coating the plate with silica gel F254.
Determine the weight per ml of the syrup (2.4.29), and calculate
the percentage content of C8H9NO2, weight in volume. Mobile phase. A mixture of 65 volumes of chloroform,
25 volumes of acetone and 10 volumes of toluene.
Storage. Store protected from light and moisture.
Test solution (a). Transfer a quantity of the powdered tablets
containing 1 g of Paracetamol to a ground-glass stoppered
15-ml centrifuge tube, add 5 ml of peroxide-free ether, shake
Paracetamol Tablets mechanically for 30 minutes, centrifuge at 1000 rpm for
Acetaminophen Tablets 15 minutes or until a clear supernatant liquid is obtained and
use the supernatant liquid.
Paracetamol Tablets contain not less than 95.0 per cent and
not more than 105.0 per cent of the stated amount of Test solution (b). Dilute 1 ml of the test solution to 10 ml with
paracetamol, C8H9NO2. ethanol (95 per cent).
Reference solution (a). A 0.005 per cent w/v solution of
Identification
4-chloroacetanilide in ethanol (95 per cent).
Extract a quantity of the powdered tablets containing 0.5 g of
Reference solution (b). Dissolve 0.25 g of 4-chloroacetanilide
Paracetamol with 20 ml of acetone, filter, evaporate the filtrate
and 0.1 g of the substance under examination in sufficient
to dryness and dry at 105º. The residue complies with the
ethanol (95 per cent) to produce 100 ml.
following tests.
Apply to the plate 200 µl of test solution (a) and 40 µl of each
A. Determine by infrared absorption spectrophotometry (2.4.6).
of test solution (b) and reference solutions (a) and (b). After
Compare the spectrum with that obtained with paracetamol
development, dry the plate in a current of warm air and examine
RS or with the reference spectrum of paracetamol.
in ultraviolet light at 254 nm. Any spot corresponding to
B. Boil 0.1 g in 1 ml of hydrochloric acid for 3 minutes, add 4-chloroacetanilide in the chromatogram obtained with test
10 ml of water and cool; no precipitate is produced. Add solution (a) is not more intense than the spot in the
0.05 ml of 0.0167 M potassium dichromate; a violet colour chromatogram obtained with reference solution (a). Any other
develops which does not turn red. secondary spot in the chromatogram obtained with test
solution (b) is not more intense than the spot in the
Tests
chromatogram obtained with reference solution (a). The
4-Aminophenol. Determine by liquid chromatography (2.4.14). chromatogram obtained with reference solution (b) shows two

902
IP 2007 LIQUID PARAFFIN

clearly separated spots, the spot corresponding to paracetamol Liquid Paraffin


having the lower Rf value.
White Mineral Oil; Liquid Petrolatum
Dissolution (2.5.2).
Liquid Paraffin is a purified mixture of liquid hydrocarbons
Apparatus. No 1
obtained from petroleum to which not more than 10 ppm of
Medium. 900 ml of phosphate buffer pH 5.8
tocopherol or of butylated hydroxytoluene may be added.
Speed and time. 50 rpm and 30 minutes.
Description. A transparent, colourless, oily liquid, free from
Withdraw a suitable volume of the medium and filter and dilute fluorescence by daylight; odourless or almost odourless.
a suitable volume of the filtrate with the same solvent. Measure
the absorbance of the resulting solution at the maximum at Tests
about 243 nm (2.4.7). Similarly measure the absorbance of a
solution of known concentration of paracetamol RS. Calculate Weight per ml (2.4.29). 0.860 g to 0.904 g.
the content of C8H9NO2. Dynamic viscosity (2.4.28). 110 mPas to 230 mPas, determined
at 20º ± 1º by Method B.
D. Not less than 80 per cent of the stated amount of C8H9NO2.
Acidity or alkalinity. To 10.0 g add 20 ml of boiling water, heat
Other tests. Complies with the tests stated under Tablets.
in a water-bath for 5 minutes, shake vigorously for 1 minute,
Assay. Weigh and powder 20 tablets. Weigh accurately a cool, allow to separate and filter the aqueous layer. To 10 ml of
quantity of the powder containing about 0.15 g of Paracetamol, the filtrate add 0.1 ml of phenolphthalein solution. The
add 50 ml of 0.1 M sodium hydroxide, dilute with 100 ml of solution is colourless and not more than 0.1 ml of 0.1 M sodium
water, shake for 15 minutes and add sufficient water to produce hydroxide is required to change the colour of the solution to
200.0 ml. Mix, filter and dilute 10.0 ml of the filtrate to 100.0 ml pink.
with water. To 10.0 ml of the resulting solution add 10 ml of Light absorption. When examined in the range 240 nm to
0.1 M sodium hydroxide, dilute to 100.0 ml with water and 280 nm (2.4.7), a 2.0 per cent w/v solution in 2,2,4-trimethyl-
mix. Measure the absorbance of the resulting solution at the pentane shows an absorption of not more than 0.1.
maximum at about 257 nm (2.4.7). Calculate the content of
C8H9NO2 taking 715 as the specific absorbance at 257 nm. Readily carbonisable substances. Place 5 ml in a dry, heat-
resistant glass-stoppered test-tube (125 mm x 18 mm)
Storage. Store protected from light and moisture. previously rinsed with chromic acid solution, then with water
and dried. Add 5 ml of nitrogen-free sulphuric acid
(containing 94.5 per cent to 95.5 per cent w/w of H2SO4), insert
the stopper and shake as vigorously as possible in the
Hard Paraffin longitudinal direction of the tube for 5 seconds. Loosen the
stopper, immediately place the tube in a bath of boiling water,
Hard Paraffin is a purified mixture of solid hydrocarbons supporting it so as to prevent contact of the tube with the
obtained from petroleum or from shale oil. bottom or side of the bath and heat for 10 minutes. At the end
Description. A white or colourless, translucent mass, of the second, fourth, sixth, and eighth minutes, remove the
frequently showing a crystalline structure; odourless even tube from the bath and shake as vigorously as possible in the
when freshly cut; slightly greasy to the touch. Burns with a longitudinal direction of the tube for 5 seconds. At the end of
luminous flame. When melted, the liquid is free from 10 minutes from the time the tube was placed in the bath
fluorescence by daylight. remove the tube and allow to stand for 10 minutes. The lower
acid layer is not more intensely coloured than a mixture of 3 ml
Tests of FCS, 1.5 ml of CCS and 0.5 ml of CSS (2.4.1), overlaid with
5 ml of liquid paraffin. If the sulphuric acid remains dispersed
Acidity or alkalinity. To 10.0 g add 20 ml of boiling water, heat
in the molten paraffin, the colour of the emulsion is not darker
in a water-bath for 5 minutes, shake vigorously for 1 minute,
than that of the standard mixture when shaken vigorously.
cool, allow to separate and filter the aqueous layer. To 10 ml of
the filtrate add 0.1 ml of phenolphthalein solution. The Solid paraffins. Place a suitable quantity, previously dried by
solution is colourless and not more than 0.1 ml of 0.1 M sodium heating at 100º for 2 hours and cooled in a desiccator over
hydroxide is required to change the colour of the solution to sulphuric acid, in a glass cylindrical vessel having an internal
pink. diameter of approximately 25 mm. Close the vessel and immerse
in a mixture of ice and water; after 4 hours the liquid is
Congealing range (2.4.10). 50º to 65º.
sufficiently clear that a black line, 0.5 mm in width, held vertically
Sulphated ash (2.3.18). Not more than 0.1 per cent. behind the vessel is easily seen.

903
LIGHT LIQUID PARAFFIN IP 2007

Sulphur compounds. Mix 4 ml with 2 ml of ethanol (95 per dispersed in the molten paraffin, the colour of the emulsion is
cent), and 2 drops of a clear, saturated solution of lead not darker than that of the standard mixture when shaken
monoxide in sodium hydroxide solution and heat at 70º for vigorously.
10 minutes with frequent shaking; the mixture remains Solid paraffins. Place a suitable quantity, previously dried by
colourless. heating at 100º for 2 hours and cooled in a desiccator over
Storage. Store protected from light. sulphuric acid, in a glass cylindrical vessel having an internal
diameter of approximately 25 mm. Close the vessel and immerse
in a mixture of ice and water; after 4 hours the liquid is
sufficiently clear that a black line, 0.5 mm in width, held vertically
Light Liquid Paraffin behind the vessel is easily seen.
Light Mineral Oil; Light Liquid Petrolatum Sulphur compounds. Mix 4 ml with 2 ml of ethanol (95 per
cent), and 2 drops of a clear, saturated solution of lead
Light Liquid Paraffin is a purified mixture of liquid saturated
monoxide in sodium hydroxide solution and heat at 70º for
hydrocarbons obtained from petroleum. It may contain a
10 minutes with frequent shaking; the mixture remains
suitable stabiliser.
colourless.
Description. A transparent, colourless, oily liquid, free from
Storage. Store protected from light.
fluorescence by daylight; almost odourless when cold.

Tests
Weight per ml (2.4.29). 0.820 g to 0.880 g. Liquid Paraffin Emulsion
Dynamic viscosity (2.4.28). 25 mPa s to 80 mPa s, determined Liquid Paraffin Oral Emulsion
at 20º ± 1º by Method B. Liquid Paraffin Emulsion is an oral emulsion of Liquid Paraffin
Acidity or alkalinity. To 10.0 g add 20 ml of boiling water, heat in Purified Water.
in a water-bath for 5 minutes, shake vigorously for 1 minute, Liquid Paraffin Emulsion contains not less than 44.0 per cent
cool, allow to separate and filter the aqueous layer. To 10 ml of and not more than 49.0 per cent w/w of liquid paraffin.
the filtrate add 0.1 ml of phenolphthalein solution. The
solution is colourless and not more than 0.1 ml of 0.1 M sodium Tests
hydroxide is required to change the colour of the solution to
pink. Other tests. Complies with the tests stated under Oral Liquids.
Light absorption. When examined in the range 240 nm to 280 Assay. Weigh accurately about 5.0 g, add 10 ml of water, extract
nm (2.4.7), a 2.0 per cent w/v solution in 2,2,4-trimethylpentane with two quantities, each of 40 ml, of a mixture of 2 volumes of
shows an absorption of not more than 0.1. ethanol (95 per cent), 3 volumes of light petroleum (40º to
60º) and 3 volumes of ether and then with 30 ml of a mixture of
Readily carbonisable substances. Place 5 ml in a dry, heat- equal volumes of light petroleum (40º to 60º) and ether. Wash
resistant glass-stoppered test-tube (125 mm x 18 mm) the combined extracts with 15 ml of 0.5 M sodium hydroxide
previously rinsed with chromic acid solution, then with water and then with 15 ml of water, evaporate the solvent, add 5 ml
and dried. Add 5 ml of nitrogen-free sulphuric acid of acetone and evaporate again. Repeat the addition and
(containing 94.5 per cent to 95.5 per cent w/w of H2SO4), insert evaporation of acetone until the residue is free from water,
the stopper and shake as vigorously as possible in the dry at 105º for 15 minutes and weigh.
longitudinal direction of the tube for 5 seconds. Loosen the
stopper, immediately place the tube in a bath of boiling water, Storage. Store protected from moisture.
supporting it so as to prevent contact of the tube with the
bottom or side of the bath and heat for 10 minutes. At the end
of the second, fourth, sixth, and eighth minutes, remove the
tube from the bath and shake as vigorously ssas possible in
White Soft Paraffin
the longitudinal direction of the tube for 5 seconds. At the White Petroleum Jelly
end of 10 minutes from the time the tube was placed in the
White Soft Paraffin is a purified, semi-solid mixture of
bath remove the tube and allow to stand for 10 minutes. The
hydrocarbons obtained from petroleum and bleached.
lower acid layer is not more intensely coloured than a mixture
of 3 ml of FCS, 1.5 ml of CCS and 0.5 ml of CSS (2.4.1), overlaid Description. A white, translucent, soft unctuous mass,
with 5 ml of liquid paraffin. If the sulphuric acid remains retaining these characteristics on storage and when melted

904
IP 2007 YELLOW SOFT PARAFFIN

and allowed to cool without stirring; not more than slightly and read the total penetration from the scale. Make three or
fluorescent by daylight, even melted; odourless when rubbed more trials, each so spaced that there is no overlapping of the
on the skin. areas of penetration. Where the penetration exceeds 20 mm,
use a separate container of the test substance for each trial.
Tests Read the penetration to the nearest 0.1 mm. Calculate the
average of the three or more readings and conduct further
Melting range (2.4.21). 38º to 56º, determined by Method IV. trials to a total of 10 if the individual results differ from the
Acidity or alkalinity. To 10.0 g add 20 ml of boiling water, heat average by more than ± 3 per cent. The final average of the
in a water-bath for 5 minutes, shake vigorously for 1 minute, trials is not less than 10.0 mm and not more than 30.0 mm
cool, allow to separate and filter the aqueous layer. To 10 ml of indicating a consistency value between 100 and 300.
the filtrate add 0.1 ml of phenolphthalein solution. The Sulphated ash (2.3.18). Not more than 0.1 per cent.
solution is colourless and not more than 0.1 ml of 0.1 M sodium
hydroxide is required to change the colour of the solution to Storage. Store protected from light and moisture.
pink.
Light absorption (2.4.7). Absorbance of a 0.05 per cent w/v
solution in 2,2,4-trimethylpentane at about 290 nm, not more Yellow Soft Paraffin
than 0.5.
Yellow Petroleum Jelly
Fixed oils, fats and resin. Digest 10 g with 50 ml of sodium
hydroxide solution at 100º for 30 minutes and allow the Yellow Soft Paraffin is a purified, semi-solid mixture of
aqueous layer to separate. On acidifying the aqueous layer hydrocarbons obtained from petroleum.
with dilute sulphuric acid, no precipitate or oily matter is Description. A pale yellow to yellow, translucent, soft
produced. unctuous mass, retaining these characteristics on storage and
Foreign organic matter. Volatilises when heated, without when melted and allowed to cool without stirring; not more
emitting an acrid odour. than slightly fluorescent by daylight, even melted; odourless
when rubbed on the skin.
Consistency. 100 to 300, determined by the following method.
Apparatus. The apparatus is essentially in agreement with IS Tests
4887:1968 and comprises a penetrometer fitted with a polished Melting range (2.4.21). 38º to 56º, determined by Method IV.
cone-shaped metal plunger weighing 150 g having a detachable
steel tip of the following dimensions. The tip of the cone has Light absorption (2.4.7). Absorbance of a 0.05 per cent w/v
an angle of 30º, the point being truncated to a diameter of solution in 2,2,4-trimethylpentane at about 290 nm, not more
0.38 ± 0.08 mm, the base of the tip is 8.38 ± 0.13 mm in diameter than 0.75.
and the length of the tip is 15 ± 0.25 mm. The remaining portion Acidity or alkalinity. To 10.0 g add 20 ml of boiling water, heat
of the cone has an angle of 90º, is 28 to 29 mm in height, and in a water-bath for 5 minutes, shake vigorously for 1 minute,
has a maximum diameter of 65.1 mm at the base. The containers cool, allow to separate and filter the aqueous layer. To 10 ml of
of the test are flat-bottomed metal or glass cylinders that are the filtrate add 0.1 ml of phenolphthalein solution. The
102 ± 6 mm in diameter and not less than 60 mm in height. solution is colourless and not more than 0.1 ml of 0.1 M sodium
Procedure. Melt a sufficient quantity at a temperature below hydroxide is required to change the colour of the solution to
85º and pour into one or more of the containers filling to within pink.
6 mm of the rim. Cool to 25º± 2.5º over a period of not less than Fixed oils, fats and resin. Digest 10 g with 50 ml of sodium
16 hours, protected from drafts. Two hours before the test, hydroxide solution at 100º for 30 minutes and allow the
place the containers in a water-bath at 25º ± 0.5º. If the room aqueous layer to separate. On acidifying the aqueous layer
temperature is below 23.5º or above 26.5º, adjust the with dilute sulphuric acid, no precipitate or oily matter is
temperature of the cone to 25º ± 0.5º by placing it in a water- produced.
bath.
Foreign organic matter. Volatilises when heated, without
Without disturbing the surface of the substance under emitting an acrid odour.
examination, place the container on the penetrometer table,
Consistency. 100 to 300, determined by the following method.
and lower the cone until the tip just touches the top surface of
the test substance at a spot 25 mm to 38 mm from the edge of Apparatus. The apparatus is essentially in agreement with IS
the container. Adjust the zero setting and quickly release the 4887.1968 and comprises a penetrometer fitted with a polished
plunger, then hold it free for 5 seconds. Secure the plunger cone-shaped metal plunger weighing 150 g having a detachable

905
PARAFFIN OINTMENT IP 2007

steel tip of the following dimensions. The tip of the cone has Paraldehyde
an angle of 30º, the point being truncated to a diameter of
0.38 ± 0.08 mm, the base of the tip is 8.38 ± 0.13 mm in diameter H3C O CH3
and the length of the tip is 15 ± 0.25 mm. The remaining portion
of the cone has an angle of 90º, is 28 to 29 mm in height, and O O
has a maximum diameter of 65.1 mm at the base. The containers
of the test are flat-bottomed metal or glass cylinders that are CH3
102 ± 6 mm in diameter and not less than 60 mm in height.
C6H12O3 Mol. Wt. 132.7
Procedure. Melt a sufficient quantity at a temperature below
Paraldehyde is 2,4,6-trimethyl-1,3,5-trioxane, the cyclic trimer
85º and pour into one or more of the containers filling to within
of acetaldehyde. It may contain a suitable amount of
6 mm of the rim. Cool to 25º± 2.5º over a period of not less than
antioxidant.
16 hours, protected from drafts. Two hours before the test,
place the containers in a water-bath at 25º ± 0.5º. If the room Description. A colourless or slightly yellow, transparent liquid;
temperature is below 23.5º or above 26.5º, adjust the odour, strong and characteristic. Solidifies at low temperature
temperature of the cone to 25º ± 0.5º by placing it in a water- to form a crystalline mass.
bath.
Identification
Without disturbing the surface of the substance under
examination, place the container on the penetrometer table, A. Heat 5 ml with 0.1 ml of 1 M sulphuric acid; acetaldehyde,
and lower the cone until the tip just touches the top surface of recognisable by its odour, is evolved.
the test substance at a spot 25 mm to 38 mm from the edge of
B. To 5 ml of a 10 per cent v/v solution add 5 ml of ammoniacal
the container. Adjust the zero setting and quickly release the
silver nitrate solution in a test-tube and heat on a water-bath;
plunger, then hold it free for 5 seconds. Secure the plunger
metallic silver is deposited as a mirror on the sides of the tube.
and read the total penetration from the scale. Make three or
more trials, each so spaced that there is no overlapping of the C. A 10 per cent w/v solution in carbon dioxide-free water is
areas of penetration. Where the penetration exceeds 20 mm, clear (2.4.1), but becomes turbid on warming.
use a separate container of the test substance for each trial.
Read the penetration to the nearest 0.1 mm. Calculate the Tests
average of the three or more readings and conduct further Congealing range (2.4.10). 10º to 13º.
trials to a total of 10 if the individual results differ from the
average by more than ± 3 per cent. The final average of the Distillation range (2.4.8). Not more than 10 per cent distils
trials is not less than 10.0 mm and not more than 30.0 mm below 123º and not less than 95 per cent distils below 126º.
indicating a consistency value between 100 and 300. Refractive index (2.4.27). 1.403 to 1.406.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Relative density (2.4.29). 0.991 to 0.996.
Storage. Store protected from light and moisture. Acetaldehyde. Shake 5 ml with a mixture of 5 ml of ethanol
(60 per cent), 5 ml of hydroxylamine hydrochloride reagent
in ethanol (60 per cent) and 2 drops of methyl orange solution
and titrate with 0.5 M sodium hydroxide to full yellow colour;
Paraffin Ointment not more than 0.8 ml of 0.5 M sodium hydroxide is required.
White Beeswax 20 g Acidity. Mix 5 ml with 45 ml of carbon dioxide-free water and
Hard Paraffin 30 g titrate with 0.1 M sodium hydroxide using phenolphthalein
Cetostearyl Alcohol 50 g solution as indicator; not more than 1.5 ml is required.
White Soft Paraffin* 900 g Chlorides. To 5 ml of a 1 per cent v/v solution add one drop of
*May be replaced by Yellow Soft Paraffin if other medicaments nitric acid and three drops of silver nitrate solution; no
to be incorporated are coloured. opalescence is produced immediately.
Mix the ingredients, heat gently with stirring until Sulphates. To 5 ml of a 1 per cent v/v solution add one drop of
homogeneous and stir until cold. hydrochloric acid and three drops of barium chloride
solution; no turbidity is produced.
Tests Peroxides. In a stoppered vessel, dissolve 5 ml in sufficient of
Paraffin Ointment complies with the tests stated under recently boiled and cooled water to produce 50 ml, add 5 ml of
Ointments. dilute sulphuric acid and 10 ml of potassium iodide solution.

906
IP 2007 PENICILLAMINE

Close the flask and set aside in the dark for 15 minutes. Titrate and expose to iodine vapour for 5 to 10 minutes. The principal
with 0.1 M sodium thiosulphate using starch solution as spot in the chromatogram obtained with the test solution
indicator; set aside for 5 minutes and, if necessary, complete corresponds to that in the chromatogram obtained with the
the titration. Not more than 2.0 ml of 0.1 M sodium reference solution.
thiosulphate is required. B. Dissolve 0.5 g in a mixture of 0.5 ml of hydrochloric acid
Non-volatile matter. Heat 5 ml in a small dish on a water-bath and 4 ml of warm acetone, cool in ice and scratch the inside of
and dry at 105º for 1 hour; the residue weighs not more than 3 the tube with a glass rod to initiate crystallisation; a white
mg (0.06 per cent w/v). precipitate is produced. Filter under vacuum, wash the
precipitate with acetone and dry with suction. A 1 per cent
Storage. Store protected from moisture, in complete darkness
w/v solution of the dried material is dextrorotatory.
and at a temperature of 8º to 15º. If solidified, the whole of the
contents of the container should be liquified by warming C. To 4 ml of a 1 per cent w/v solution add 2 ml of
before use. phosphotungstic acid solution and heat nearly to boiling; a
blue colour is produced.
NOTE — Do not use Paraldehyde if it has a brownish colour
or an odour of acetic acid. Avoid contact with rubber and D. In the test for Penicillamine disulphide, the principal peak
plastics. in the chromatogram obtained with test solution (b)
Labelling. The label states (1) the nature and the proportion corresponds to the peak due to penicillamine in the
of any antioxidant added; (2) that it may decompose on chromatogram obtained with reference solution (a).
standing to form potentially harmful substances. Tests
Appearance of solution. A 10.0 per cent w/v solution in carbon
dioxide-free water (solution A) is clear (2.4.1), and not more
Penicillamine intensely coloured than degree 6 of the appropriate range of
reference solutions (2.4.1).
D-Penicillamine
pH (2.4.24). 4.5 to 5.5, determined in a 1.0 per cent w/v solution.
CH3 O Specific optical rotation (2.4.22). –61.0º to –65.0º, determined
H3 C
OH in a 5.0 per cent w/v solution in 1 M sodium hydroxide.
HS
H NH2 Heavy metals (2.3.13). 10 ml of solution A, complies with the
limit test for heavy metals, Method D (20 ppm). Use lead
C5H11NO2S Mol. Wt. 149.2
standard solution (2 ppm Pb) to prepare the standard.
Penicillamine is 3-mercapto-D-valine.
Mercuric salts. Determine by atomic absorption
Penicillamine contains not less than 98.0 per cent and not spectrophotometry (2.4.2), using a solution prepared in the
more than 101.0 per cent of C5H11NO2S, calculated on the dried following manner. To 1.0 g of the substance under examination
basis. add 10 ml of water and 0.15 ml of perchloric acid and swirl
Description. A white or almost white, crystalline powder. until dissolution is complete. Add 1 ml of ammonium
pyrrolidinedithiocarbamate solution that has been washed
Identification three times immediately before use, each time with an equal
volume of 4-methyl-2-pentanone. Mix, add 2 ml of 4-methyl-
Test A may be omitted if test B, C and D carried out. Test D 2-pentanone, shake for 1 minute, dilute to 25 ml with water,
may be omitted if test A, B and C are carried out. allow the layers to separate and use the 4-methyl-2-pentanone
A. Determine by thin layer chromatography (2.4.17), coating layer. Measure the absorbance at 254 nm using a mercury
the plate with silica gel G. hollow-cathode lamp and an air-acetylene flame and setting
the zero using a 4-methyl-2-pentanone layer obtained by
Mobile phase. A mixture of 40 volumes of 1-butanol,
repeating the procedure described above but omitting the
10 volumes of glacial acetic acid and 10 volumes of water.
substance under examination. For the standard solution
Test solution. Dissolve 0.25 g of the substance under dissolve 0.108 g of yellow mercuric oxide in the minimum
examination in 100 ml of water. volume of 2 M hydrochloric acid, add sufficient water to
Reference solution. A 0.25 per cent w/v solution of produce 1000.0 ml and treat suitable volumes in the same manner
penicillamine RS in water. as the solution of the substance under examination (10 ppm).
Apply to the plate 2 µl of each solution. Allow the mobile Penicillamine disulphide. Determine by liquid
phase to rise 10 cm. Dry the plate at 105º for 5 to 10 minutes chromatography (2.4.14).

907
PENICILLAMINE IP 2007

Test solution (a). Dissolve 40 mg of the substance under ether and shake vigorously for 1 minute. Repeat the extraction
examination in 5 ml of the mobile phase, add 1 ml of a 0.0025 and combine the ether layers. Add 8 ml of phosphate buffer
per cent w/v solution of sulphanilamide (internal standard) pH 2.5, shake for 1 minute, allow to settle and separate the
in the mobile phase and dilute to 10 ml with the mobile phase. ether layer quantitatively, taking care to eliminate the aqueous
phase completely. (Penicillin is unstable at pH 2.5; carry out
Test solution (b). Dissolve 40 mg of the substance under
the operations at this pH within 6 to 7 minutes). Add 8 ml of
examination in the mobile phase and dilute to 10 ml with the
phosphate buffer pH 6.0, shake for 5 minutes, allow to settle,
same solvent.
separate the aqueous layer and check that the pH is 6.0. For
Reference solution (a). A 0.4 per cent w/v solution of solution (2) add 20 µl of penicillinase solution to 2 ml of
penicillamine RS in the mobile phase. solution (1) and incubate at 37º for 1 hour. For solution (3)
dissolve 5 mg of benzylpenicillin sodium in 500 ml of
Reference solution (b). Add 1 ml of test solution (a) to 1 ml of
phosphate buffer pH 6.0 and dilute 0.25 ml of this solution to
a 0.04 per cent w/v solution of penicillamine disulphide RS in
200 ml with phosphate buffer pH 2.5. Carry out the extraction
the mobile phase and dilute to 10 ml with the mobile phase.
procedure described under solution (1) using 8 ml of this
Chromatographic system solution and beginning at the words “add 8 ml of ether...”. For
– a stainless steel column 25 cm x 5 mm, packed with solution (4) add 20 ml of penicillinase solution to 2 ml of
octylsilane bonded to porous silica (5 to 10 µm), solution (3) and incubate at 37º for 1 hour. Prepare solution (5)
– mobile phase: an equal volume of 0.2 per cent w/v in the same manner as solution (1) but omitting the substance
solution of methanesulphonic acid and 0.01 per cent under examination.
w/v solution of disodium edetate, Maintain the dishes at 30º for at least 24 hours. Measure the
– flow rate. 2 ml per minute, diameters of the zones of inhibition to within 0.1 mm. The test
– spectrophotometer set at 220 nm, is not valid unless solution (3) gives a clear zone of inhibition
– a 20 µl loop injector. and solutions (4) and (5) give no zones of inhibition. If solution
In the chromatogram obtained with test solution (a) the ratio (1) gives a zone of inhibition it is caused by penicillin provided
of the area of any peak corresponding to penicillamine solution (2) gives no zone of inhibition. If this is the case, the
disulphide to the area of the peak due to the internal standard average diameter of the zones of inhibition given by solution
is not greater than the corresponding ratio in the chromatogram (1) for the five Petri dishes is less than that given by solution
obtained with reference solution (b). (3) (0.1 ppm).
Penicillin. Carry out the following procedure in a penicillin- Nutrient medium
free atmosphere and with equipment reserved for the test. Peptone 5 g
Sterilise the equipment at 180º for 3 hours and the buffer
Yeast extract 1.5 g
solutions at 121º for 20 minutes before use.
Meat extract 1.5 g
Liquefy a suitable nutrient medium such as that described Sodium chloride 3.5 g
below and inoculate at a suitable temperature with a culture of
Agar 15 g
Micrococcus flavus (ATCC 9341) to give 5 x 104 micro-
organisms per ml or a quantity necessary to obtain the required Distilled water 1000 ml
sensitivity and formation of clearly defined inhibition zones Adjust the pH to 6.0
of suitable diameter. Immediately pour the inoculated medium Penillic acid. Absorbance of a 0.2 per cent w/v solution at
into five Petri dishes (10 cm in diameter) to give uniform layers about 268 nm, not more than 0.07 (2.4.7) (about 0.5 per cent).
2 to 5 mm in depth. Alternatively, the medium may consist of
two layers, only the upper layer being inoculated. Store the Sulphated ash (2.3.18). Not more than 0.1 per cent.
dishes so that no appreciable growth or death of micro- Loss on drying (2.4.19). Not more than 0.5 per cent, determined
organisms occurs before use and so that the surface of the on 1.0 g by drying in an oven at 60º over phosphorus pentoxide
medium is dry at the time of use. In each dish, place five at a pressure not exceeding 0.7 kPa.
stainless steel hollow cylinders (6 mm in diameter) on the
Assay. Dissolve 0.1 g in 30 ml of anhydrous glacial acetic
surface of the medium evenly spaced on a circle with a radius
acid. Titrate with 0.1 M perchloric acid, determining the end-
of about 25 mm and concentric with the dish. For each dish,
point potentiometrically (2.4.25). Carry out a blank titration.
place in separate cylinders 0.15 ml of each of the following
five solutions. 1 ml of 0.1 M perchloric acid is equivalent to 0.01492 g of
C5H11NO2S.
For solution (1) dissolve 1.0 g of the substance under
examination in 8 ml of phosphate buffer pH 2.5, add 8 ml of Storage. Store protected from moisture.

908
IP 2007 DILUTED PENTAERYTHRITOL TETRANITRATE

Penicillamine Tablets In the chromatogram obtained with the test solution the area
of any peak corresponding to penicillamine disulphide is not
D-Penicillamine Tablets greater than the area of the principal peak in the chromatogram
Penicillamine Tablets contain not less than 95.0 per cent and obtained with the reference solution (1.0 per cent).
not more than 105.0 per cent of the stated amount of Other tests. Complies with the tests stated under Tablets.
penicillamine, C5H11NO2S. The tablets are coated.
Assay. Weigh and powder 20 tablets. Weigh accurately a
Identification quantity of the powder containing 0.1 g of Penicillamine,
dissolve as completely as possible in 50 ml of water and filter.
A. Shake a quantity of the powdered tablets containing 20 mg Add to the filtrate 5 ml of 1 M sodium hydroxide and 0.2 ml of
of Penicillamine with 4 ml of water and filter. Add to the filtrate a 0.1 per cent w/v solution of dithizone in ethanol (95 per
2 ml of phosphotungstic acid solution and allow to stand for cent) and titrate with 0.02 M mercuric nitrate.
5 minutes; a blue colour is produced.
1 ml of 0.02 M mercuric nitrate is equivalent to 0.005968 g of
B. Dissolve a quantity of the powdered tablets containing C5H11NO2S.
10 mg of Penicillamine in 5 ml of water and add 0.3 ml of 5 M
Storage. Store protected from moisture.
sodium hydroxide and 20 mg of ninhydrin; an intense blue or
violet-blue colour is produced immediately.

Tests
Mercuric salts. Disperse a quantity of the powdered tablets
Diluted Pentaerythritol Tetranitrate
containing 1 g of Penicillamine in 10 ml of water in a stoppered
flask, add 0.2 ml of 9 M perchloric acid and swirl to dissolve. O2 N NO2
O O
Add 1 ml of ammonium pyrrolidinedithiocarbamate solution,
mix, add 2 ml of 4-methyl-2-pentanone, shake for 1 minute and
O2N O O
add sufficient water to produce 25 ml. Determine by atomic NO2
absorption spectrophotometry (2.4.2), using a mercury hollow-
cathode lamp and an air-acetylene flame and setting the zero C5H8N4O12 Mol. Wt. 316.1
using a 4-methyl-2-pentanone layer obtained by repeating
the procedure described above but omitting the substance Diluted Pentaerythritol Tetranitrate is a dry mixture of 2,2-
under examination, measuring at 254 nm. Use mercury solution bis(hydroxymethyl)- propane1,3-diol tetranitrate with
AAS, suitably diluted with water, for the standard solutions, Lactose or Mannitol or a mixture of Lactose and Starch or
adjusted to contain the same concentrations of 9 M perchloric any other suitable inert excipients which permit safe
acid, ammonium pyrrolidinedithiocarbamate solution and handling.
4-methyl-2-pentanone as the solution under examination Diluted Pentaerythritol Tetranitrate contains not less than
(40 ppm). 95.0 per cent and not more than 105.0 per cent of the stated
Penicillamine disulphide. Determine by liquid amount of pentaerythritol tetranitrate, C5H8N4O12.
chromatography (2.4.14). Description. A white or almost white, powder; odour, faint
Test solution. Shake quantity of the powdered tablets and mild.
containing 40 mg of Penicillamine with 10 ml of the mobile
phase, filter and use the filtrate. Identification
Reference solution. A 0.004 per cent w/v solution of A. Transfer a quantity containing 10 mg of pentaerythritol
penicillamine disulphide RS in the mobile phase. tetranitrate to a medium porosity sintered-glass filter, add 5 ml
Chromatographic system of dry acetone and collect the filtrate. Repeat with two further
– a stainless steel column 25 cm x 5 mm, packed with quantities, each of 5 ml, of dry acetone and evaporate the
octylsilane bonded to porous silica (5 to 10 µm), combined filtrate at a temperature not exceeding 60º, with the
– mobile phase: an equal volume of 0.2 per cent w/v aid of a gentle current of air, and dry the residue at 60º for
solution of methanesulphonic acid and 0.01 per cent 4 hours; the residue melts at 138º to 142º (2.4.21).
w/v solution of disodium edetate, B. Suspend 10 mg of the residue obtained in test A in a mixture
– flow rate. 2 ml per minute, of 2 ml of sulphuric acid and 1 ml of water; cool and carefully
– spectrophotometer set at 220 nm, overlay with 3 ml of ferrous sulphate solution; a reddish brown
– a 20 µl loop injector. colour is produced at the interface of the two liquids.

909
PENTAERYTHRITOL TETRANITRATE TABLETS IP 2007

Tests of dry acetone and collect the filtrate. Repeat with two further
quantities, each of 5 ml, of dry acetone and evaporate the
Heavy metals (2.3.13). 2.0 g complies with limit test for heavy combined filtrate at a temperature not exceeding 60º, with the
metals, Method B (10 ppm). aid of a gentle current of air, and dry the residue at 60º for
Assay. Weigh accurately a quantity containing about 50 mg 4 hours; the residue melts at 138º to 142º (2.4.21).
of pentaerythritol tetranitrate and transfer to a 100-ml
volumetric flask with the aid of about 30 ml of acetone. Add Tests
sufficient acetone to produce 50 ml and warm on a water-bath
Uniformity of content. (For tablets containing 10 mg or less)
at a temperature not exceeding 60º and boil gently, with
— Comply with the test stated under Tablets.
occasional swirling, for 5 minutes. Cool, dilute to volume with
acetone and mix. Transfer a portion of the mixture to a glass- Crush one tablet and transfer to a 50-ml volumetric flask with
stoppered centrifuge tube and centrifuge at 1500 rpm for the aid of 15 ml of acetone. Add sufficient acetone to produce
5 minutes. Transfer 1.0 ml of the supernatant solution to a 25 ml, heat the mixture on a water-bath at a temperature not
100-ml volumetric flask and evaporate at 35º with the aid of a exceeding 60º and boil gently, with occasional swirling, for
current of air to dryness. To the residue add 1.0 ml of glacial 5 minutes. Cool, dilute to volume with acetone and mix.
acetic acid and swirl to dissolve. Add 2 ml of Transfer a portion of the mixture to a glass-stoppered
phenoldisulphonic acid solution, mix and allow to stand for centrifuge tube and centrifuge at 1500 rpm for 5 minutes.
5 minutes. Add 25 ml of water and 10 ml of strong ammonia Transfer 2.5 ml of the supernatant solution to a 100-ml
solution, cool, dilute to volume with water and mix. Measure volumetric flask and evaporate at 35º with the aid of a current
the absorbance of the resulting solution at the maximum at of air to dryness. To the residue add 1.0 ml of glacial acetic
about 409 nm (2.4.7), using water as the blank. acid and swirl to dissolve. Add 2 ml of phenoldisulphonic
acid solution, mix and allow to stand for 5 minutes. Add 25 ml
Weigh accurately 0.13 g of potassium nitrate, previously dried
of water and 10 ml of strong ammonia solution, cool, dilute to
at 105º for 4 hours, dissolve in 3 ml of water, dilute with
volume with water and mix. Measure the absorbance of the
sufficient glacial acetic acid to produce 200.0 ml and mix
resulting solution at the maximum at about 409 nm (2.4.7),
well. Using 1.0 ml of this solution repeat the procedure
using water as the blank.
beginning at the words “Add 2 ml of phenoldisulphonic acid
solution,.......”. Weigh accurately 0.130 g of potassium nitrate, previously
dried at 105º for 4 hours, dissolve in 3 ml of water, dilute with
Calculate the content of C5H8N4O12 from the values of the
sufficient glacial acetic acid to produce 200.0 ml and mix
absorbances so obtained.
well. Using 1.0 ml of this solution repeat the procedure
1 ml of the potassium nitrate solution is equivalent to beginning at the words “Add 2 ml of phenoldisulphonic acid
0.000503 g of C5H8N4O12. solution,.......”.Calculate the content of C5H8N4O12 in the tablet.
Storage. Store protected from light and moisture. 1 ml of the potassium nitrate solution is equivalent to 0.000503
NOTE — Undiluted pentaerythritol tetranitrate is a powerful g of C5H8N4O12.
explosive. It can be exploded with percussion or excessive Other tests. Complies with the tests stated under Tablets.
heat. Great care and appropriate precautions should be
taken in handling and only exceedingly small amounts should Assay. Weigh and powder 20 tablets. Weigh accurately a
be isolated. quantity of the powder containing about 50 mg of
pentaerythritol tetranitrate and transfer to a 100-ml volumetric
Labelling. The label states the percentage content of
flask with the aid of about 30 ml of acetone. Add sufficient
pentaerythritol tetranitrate.
acetone to produce 50 ml and warm on a water-bath at a
temperature not exceeding 60º and boil gently, with occasional
swirling, for 5 minutes. Cool, dilute to volume with acetone
Pentaerythritol Tetranitrate Tablets and mix. Transfer a portion of the mixture to a glass-stoppered
centrifuge tube and centrifuge at 1500 rpm for 5 minutes.
Pentaerythritol Tetranitrate Tablets contain not less than Transfer 1.0 ml of the supernatant solution to a 100-ml
90.0 per cent and not more than 110.0 per cent of the stated volumetric flask and evaporate at 35º with the aid of a current
amount of pentaerythritol tetranitrate, C5H8N4O12. of air to dryness. To the residue add 1.0 ml of glacial acetic
acid and swirl to dissolve. Add 2 ml of phenoldisulphonic
Identification acid solution, mix and allow to stand for 5 minutes. Add 25 ml
A. Transfer a quantity containing 10 mg of pentaerythritol of water and 10 ml of strong ammonia solution, cool, dilute to
tetranitrate to a medium porosity sintered-glass filter, add 5 ml volume with water and mix. Measure the absorbance of the

910
IP 2007 PENTAMIDINE INJECTION

resulting solution at the maximum at about 409 nm (2.4.7), Tests


using water as the blank.
pH (2.4.24). 4.5 to 6.5, determined in a 5.0 per cent w/v solution.
Weigh accurately 0.13 g of potassium nitrate, previously dried
Related substances. Determine by thin-layer chromatography
at 105º for 4 hours, dissolve in 3 ml of water, dilute with
(2.4.17), coating the plate with silica gel GF254.
sufficient glacial acetic acid to produce 200.0 ml and mix
well. Using Mobile phase. The upper layer obtained by shaking together
1.0 ml of this solution repeat the procedure beginning at the 50 volumes of water, 40 volumes of 1-butanol and 10 volumes
words “Add 2 ml of phenoldisulphonic acid solution,.......”. of glacial acetic acid.
Calculate the content of C5H8N4O12 from the values of the Test solution. Dissolve 0.5 g of the substance under
absorbances so obtained. examination in 10 ml of methanol.
1 ml of the potassium nitrate solution is equivalent to Reference solution. A 0.025 per cent w/v solution of the
0.000503 g of C5H8N4O12. substance under examination in 100 ml of methanol.

Storage. Store protected from light and moisture. Activate the plate by heating at 105º for 1 hour, apply to the
plate 10 µl of each solution. After development, dry the plate
Labelling. The label states the strength in terms of the in air and examine in ultraviolet light at 254 nm. Any secondary
equivalent amount of pentaerythritol tetranitrate. spot in the chromatogram obtained with the test solution is
not more intense than the spot in the chromatogram obtained
with the reference solution.
Pentamidine Isethionate Ammonium isethionate. To 1.0 g in a test-tube (about 4 cm in
diameter) add 10 ml of water and 20 ml of 1 M sodium
O O hydroxide. Immediately attach a bung carrying a splash head
SO3H and an aspirator tube (about 5 mm in diameter). Connect the
H2N NH2 . HO
2 splash head to two test-tubes in series, each containing 20 ml
NH NH of 0.01 M sulphuric acid. Heat the tube containing the
substance under examination in a water-bath at 45º to 50º and,
C19H24N4O2,2C2H6O4S Mol. Wt. 592.7 maintaining this temperature, draw a current of air, previously
Pentamidine Isethionate is 4,4′-[pentane-1,5- passed through 1 M sulphuric acid, through the liquids in a
diylbis(oxy)]dibenzamidine di(2-hydroxyethanesulphonate). series of tubes for 3 hours at such a rate that the bubbles are
just too rapid to count. Titrate the combined solutions from
Pentamidine Isethionate contains not less than 98.5 per cent
the two absorption tubes with 0.02 M sodium hydroxide using
and not more than 101.0 per cent of C19H24N4O2, 2C2H6O4S,
methyl red-methylene blue solution as indicator; not less than
calculated on the dried basis.
36.5 ml of 0.02 M sodium hydroxide is required.
Description. A white or almost white powder or crystals;
Sulphated ash (2.3.18). Not more than 0.1 per cent.
odourless or almost odourless; hygroscopic.
Loss on drying (2.4.19). Not more than 4.0 per cent, determined
Identification on 1.0 g by drying in an oven at 105º.
A. Determine by infrared absorption spectrophotometry (2.4.6). Assay. Dissolve 0.25 g in 50 ml of dimethylformamide. Titrate
Compare the spectrum with that obtained with pentamidine with 0.1 M tetrabutylammonium hydroxide, determining the
isethionate RS or with the reference spectrum of pentamidine end-point potentiometrically (2.4.25). Carry out a blank titration.
isethionate. 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
B. When examined in the range 230 nm to 360 nm (2.4.7), a 0.02963 g of C19H24N4O2,2C2H6O4S.
0.001 per cent w/v solution in 0.01 M hydrochloric acid shows Storage. Store protected from moisture.
an absorption maximum only at about 262 nm; absorbance at
about 262 nm, about 0.46).
C. To 10 ml of a 0.05 per cent w/v solution add 1 ml of a 0.1 per Pentamidine Injection
cent w/v solution of glyoxal sodium bisulphite and 1 ml of a
solution prepared by dissolving 4 g of boric acid in a mixture Pentamidine Isethionate Injection
of 27 ml of 1 M sodium hydroxide and sufficient water to Pentamidine Injection is a sterile material consisting of
produce 100 ml. Heat on a water-bath for 10 minutes; a magenta Pentamidine Isethionate with or without buffering agents and
colour is produced. other excipients. It is filled in a sealed container.

911
PENTAZOCINE IP 2007

The injection is constituted by dissolving the contents of the not more intense than the spot in the chromatogram obtained
sealed container in the requisite amount of sterile Water for with the reference solution.
Injections, immediately before use. Ammonium isethionate. To 1.0 g in a test-tube (about 4 cm in
The constituted solution complies with the requirements for diameter) add 10 ml of water and 20 ml of 1 M sodium
Clarity of solution and Particulate matter stated under hydroxide. Immediately attach a bung carrying a splash head
Parenteral Preparations (Injections). and an aspirator tube (about 5 mm in diameter). Connect the
splash head to two test-tubes in series, each containing 20 ml
Storage. The constituted solution should be used immediately
of 0.01 M sulphuric acid. Heat the tube containing the
after preparation but, in any case, within the period
substance under examination in a water-bath at 45º to 50º and,
recommended by the manufacturer.
maintaining this temperature, draw a current of air, previously
Pentamidine Injection contains not less than 95.0 per cent and passed through 1 M sulphuric acid, through the liquids in a
not more than 105.0 per cent of the stated amount of series of tubes for 3 hours at such a rate that the bubbles are
pentamidine isethionate, C19H24N4O2, 2C2H6O4S. just too rapid to count. Titrate the combined solutions from
The contents of the sealed container comply with the the two absorption tubes with 0.02 M sodium hydroxide using
requirements stated under Parenteral Preparations methyl red-methylene blue solution as indicator; not less than
(Powders for Injection) and with the following requirements. 36.5 ml of 0.02 M sodium hydroxide is required.
Assay. Dissolve 0.25 g of the mixed contents of 10 containers
Identification in 50 ml of dimethylformamide. Titrate with 0.1 M
tetrabutylammonium hydroxide, determining the end-point
A. Determine by infrared absorption spectrophotometry (2.4.6).
potentiometrically (2.4.25). Carry out a blank titration.
Compare the spectrum with that obtained with pentamidine
isethionate RS or with the reference spectrum of pentamidine 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to
isethionate. 0.02963 g of C19H24N4O2, 2C2H6O4S.
B. When examined in the range 230 nm to 360 nm (2.4.7), a Storage. Store in single dose containers.
0.001 per cent w/v solution in 0.01 M hydrochloric acid shows
an absorption maximum only at about 262 nm; absorbance at
about 262 nm, about 0.46.
Pentazocine
C. To 10 ml of a 0.05 per cent w/v solution add 1 ml of a 0.1 per
cent w/v solution of glyoxal sodium bisulphite and 1 ml of a
solution prepared by dissolving 4 g of boric acid in a mixture HO
of 27 ml of 1 M sodium hydroxide and sufficient water to
produce 100 ml. Heat on a water-bath for 10 minutes; a magenta
colour is produced.
H3C N CH3
Tests CH3
CH3
pH (2.4.24). 4.5 to 6.5, determined in a 5.0 per cent w/v solution.
Related substances. Determine by thin-layer chromatography C19H27NO Mol. Wt. 285.4
(2.4.17), coating the plate with silica gel GF254. Pentazocine is (2RS,6RS,11RS)-6,11-dimethyl-3-(3-methylbut-
Mobile phase. The upper layer obtained by shaking together 2-enyl)-1,2,3,4,5,6-hexahydro-2,6-methano-3-benzazocin-8-ol.
50 volumes of water, 40 volumes of 1-butanol and 10 volumes Pentazocine contains not less than 98.0 per cent and not more
of glacial acetic acid. than 101.0 per cent of C19H27NO, calculated on the dried basis.
Test solution. Dissolve 0.5 g of the substance under Description. A white or pale cream powder.
examination in 10 ml of methanol.
Identification
Reference solution. A 0.025 per cent w/v solution of the
substance under examination in 100 ml of methanol. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with pentazocine
Activate the plate by heating at 105º for 1 hour, apply to the
RS or with the reference spectrum of pentazocine.
plate 10 µl of each solution. After development, dry the plate
in air and examine in ultraviolet light at 254 nm. Any secondary B. To 1 mg in a porcelain crucible add 0.5 ml of a solution of
spot in the chromatogram obtained with the test solution is sulphuric acid containing 1 per cent w/v solution of

912
IP 2007 PENTAZOCINE HYDROCHLORIDE

ammonium molybdate; an intense blue colour is produced Pentazocine Hydrochloride


which changes to bluish green, green and finally, on standing,
yellow.
HO
C. Dissolve 5 mg in 5 ml of sulphuric acid, add 0.05 ml of ferric
chloride solution and mix; a yellow colour is produced which
deepens slightly in intensity on warming. On the addition of , HCl
0.05 ml of nitric acid the yellow colour is unchanged. N CH3
H3C
Tests CH3
CH3
Light absorption (2.4.7). To 0.1 g add 20 ml of water and 10 ml
of 1 M hydrochloric acid, shake to dissolve and add sufficient C19H27NO,HCl Mol. Wt. 321.9
water to produce 100 ml. Dilute 10 ml to 100 ml with water. The
absorbance of the resulting solution at the maximum at about Pentazocine Hydrochloride is (2RS,6RS,11RS)-6,11-
278 nm, 0.67 to 0.71. dimethyl-3-(3-methylbut-2-enyl)-1,2,3,4,5,6-hexahydro-2,6-
methano-3-benzazocin-8-ol hydrochloride.
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel HF254. Pentazocine Hydrochloride contains not less than 98.0 per
cent and not more than 101.0 per cent of C19H27NO,HCl,
Mobile phase. A mixture of 94 volumes of chloroform, calculated on the dried basis.
3 volumes of 2-propylamine and 3 volumes of methanol.
Description. A white or pale cream-coloured, crystalline
Test solution. Dissolve 0.2 g of the substance under powder; odourless. The material exhibits polymorphism.
examination in 10 ml of chloroform.
Reference solution (a). A 0.02 per cent w/v solution of the Identification
substance under examination in chloroform.
A. Determine by infrared absorption spectrophotometry (2.4.6).
Reference solution (b). A 0.01 per cent w/v solution of the Compare the spectrum with that obtained with pentazocine
substance under examination in chloroform. hydrochloride RS or with the reference spectrum of
Reference solution (c). A 0.005 per cent w/v solution of the pentazocine hydrochloride.
substance under examination in chloroform. B. To 1 mg in a porcelain crucible add 0.5 ml of a solution of
Apply to the plate 10 µl of each solution. After development, sulphuric acid containing 1 per cent w/v solution of
dry the plate in air and examine in ultraviolet light at 254 nm. ammonium molybdate; an intense blue colour is produced
Heat the plate at 105º for 15 minutes, allow to cool, expose to which changes to bluish green, green and finally, on standing,
iodine vapour and re-examine in ultraviolet light at 254 nm. yellow.
Any secondary spot in the chromatogram obtained with the C. Gives reaction A of chlorides (2.3.1).
test solution is not more intense than the spot in the
chromatogram obtained with reference solution (a); not more Tests
than one such spot is more intense than the spot in the
chromatogram obtained with reference solution (b) and not pH (2.4.24). 4.0 to 6.0, determined in a 1.0 per cent w/v solution.
more than four such spots are more intense than the spot in Light absorption (2.4.7). Dissolve 0.1 g in 10 ml of 1 M
the chromatogram obtained with reference solution (c). hydrochloric acid and add sufficient water to produce
Sulphated ash (2.3.18). Not more than 0.1 per cent. 100 ml; dilute 10 ml to 100 ml with water. Absorbance of the
resulting solution at the maximum at about 278 nm, 0.59 to
Loss on drying (2.4.19). Not more than 1.0 per cent, determined 0.63.
on 1.0 g by drying in an oven at 60º at a pressure not exceeding
0.7 kPa for 4 hours. Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel HF254.
Assay. Weigh accurately about 0.6 g, dissolve in 50 ml of
anhydrous glacial acetic acid and titrate with 0.1 M perchloric Mobile phase. A mixture of 94 volumes of chloroform,
acid, using crystal violet solution as indicator. Carry out a 3 volumes of 2-propylamine and 3 volumes of methanol.
blank titration. Test solution. Dissolve 0.2 g of the substance under
1 ml of 0.1 M perchloric acid is equivalent to 0.02854 g of examination in 10 ml of chloroform.
C19H27NO. Reference solution (a). A 0.02 per cent w/v solution of the
Storage. Store protected from light and moisture. substance under examination in chloroform.

913
PENTAZOCINE TABLETS IP 2007

Reference solution (b). A 0.01 per cent w/v solution of the B. To a quantity of the powdered tablets containing 50 mg of
substance under examination in chloroform. Pentazocine Hydrochloride add 70 ml of water, shake for
Reference solution (c). A 0.005 per cent w/v solution of the 15 minutes, add sufficient water to produce 100 ml and filter.
substance under examination in chloroform. To 10 ml of the filtrate add 10 ml of 1 M sodium hydroxide and
sufficient water to produce 100 ml. When examined in the
Apply to the plate 10 µl of each solution. After development, range 230 nm to 360 nm (2.4.7), the resulting solution shows
dry the plate in air and examine in ultraviolet light at 254 nm. absorption maxima at about 238 nm and 298 nm.
Heat the plate at 105º for 15 minutes, allow to cool, expose to
C. Shake a quantity of the powdered tablets containing 25 mg
iodine vapour and re-examine in ultraviolet light at 254 nm.
of Pentazocine Hydrochloride with 5 ml of water and 0.5 ml of
Any secondary spot in the chromatogram obtained with the
2 M nitric acid for 1 minute and filter. The filtrate gives reaction
test solution is not more intense than the spot in the
A of chlorides (2.3.1).
chromatogram obtained with reference solution (a); not more
than one such spot is more intense than the spot in the
chromatogram obtained with reference solution (b) and not
Tests
more than four such spots are more intense than the spot in Related substances. Determine by thin-layer chromatography
the chromatogram obtained with reference solution (c). (2.4.17), coating the plate with silica gel HF254.
Sulphated ash (2.3.18). Not more than 0.1 per cent. Mobile phase. A mixture of 94 volumes of chloroform,
Loss on drying (2.4.19). Not more than 1.0 per cent, determined 3 volumes of 2-propylamine and 3 volumes of methanol.
on 1.0 g by drying in an oven at 100º at a pressure not exceeding Test solution. Shake a quantity of the powdered tablets
0.7 kPa. containing 0.2 g of Pentazocine Hydrochloride with 10 ml of
Assay. Weigh accurately about 0.5 g, dissolve in 50 ml of 0.1 M methanolic ammonia for 10 minutes, centrifuge and
anhydrous glacial acetic acid. Titrate with 0.1 M perchloric use the supernatant liquid.
acid, using crystal violet solution as indicator. Carry out a Reference solution (a). Dilute 1 volume of test solution to
blank titration. 100 volumes with the same solvent.
1 ml of 0.1 M perchloric acid is equivalent to 0.03219 g of Reference solution (b). Dilute 1 volume of test solution to
C19H27NO,HCl. 200 volumes with the same solvent.
Storage. Store protected from light and moisture. Reference solution (c). Dilute 1 volume of test solution to
400 volumes with the same solvent.
Apply to the plate 10 µl of each solution. After development,
dry the plate in air and examine in ultraviolet light at 254 nm.
Pentazocine Tablets Heat the plate at 105º for 15 minutes, allow to cool, expose to
Pentazocine Hydrochloride Tablets iodine vapour and re-examine in ultraviolet light at 254 nm.
Any secondary spot in the chromatogram obtained with the
Pentazocine Tablets contain not less than 92.5 per cent and test solution is not more intense than the spot in the
not more than 107.5 per cent of the stated amount of chromatogram obtained with reference solution (a), not more
pentazocine hydrochloride, C19H27NO,HCl. than one such spot is more intense than the spot in the
chromatogram obtained with reference solution (b) and not
Identification
more than four such spots are more intense than the spot in
A. Shake a quantity of the powdered tablets containing 0.1 g the chromatogram obtained with reference solution (c).
of Pentazocine Hydrochloride with 10 ml of water, filter, add Other tests. Complies with the tests stated under Tablets.
1 ml of 1 M sodium hydroxide and shake the resulting solution
with 20 ml of chloroform. Wash the chloroform extract with Assay. Weigh and powder 20 tablets. Weigh accurately a
5 ml of water, dry over anhydrous sodium sulphate and filter. quantity of the powder containing about 25 mg of Pentazocine
Evaporate the chloroform using a rotary evaporator and dry Hydrochloride, shake with 100 ml of water for 15 minutes, add
the oily residue at a temperature not exceeding 25º at a pressure 2.5 ml of 1 M hydrochloric acid and sufficient water to
of 2 kPa for 1 hour. produce 250.0 ml and filter. Measure the absorbance of the
filtrate at the maximum at about 278 nm (2.4.7). Calculate the
On the residue, determine by infrared absorption
content of C19H27NO,HCl taking 61.2 as the specific absorbance
spectrophotometry (2.4.6). Compare the spectrum with that
at 278 nm.
obtained with pentazocine RS or with the reference spectrum
of pentazocine. Storage. Store protected from light and moisture.

914
IP 2007 PENTAZOCINE INJECTION

Pentazocine Lactate Reference solution (b). A 0.01 per cent w/v solution of the
substance under examination in chloroform.
HO Reference solution (c). A 0.005 per cent w/v solution of the
substance under examination in chloroform.
Apply to the plate 25 µl of each solution. After development,
COOH
, dry the plate in air and examine in ultraviolet light at 254 nm.
H3C N CH3 Heat the plate at 105º for 15 minutes, allow to cool, expose to
H3C OH
CH3 iodine vapour and re-examine in ultraviolet light at 254 nm.
CH3 Any secondary spot in the chromatogram obtained with the
test solution is not more intense than the spot in the
C19H27NO,C3H6O3 Mol. Wt. 375.4 chromatogram obtained with reference solution (a); not more
than one such spot is more intense than the spot in the
Pentazocine Lactate is (2RS,6RS,11RS)-6,11-dimethyl-3-(3-
chromatogram obtained with reference solution (b) and not
methylbut-2-enyl)-1,2,3,4,5,6-hexahydro-2,6-methano-3-
more than four such spots are more intense than the spot in
benzazocin-8-ol lactate.
the chromatogram obtained with reference solution (c).
Pentazocine Lactate contains not less than 98.5 per cent and
Sulphated ash (2.3.18). Not more than 0.1 per cent.
not more than 101.0 per cent of C19H27NO,C3H6O3, calculated
on the dried basis. Loss on drying (2.4.19). Not more than 1.0 per cent, determined
on 1.0 g by drying in an oven at 60º at a pressure not exceeding
Description. A white or pale cream-coloured, crystalline
0.7 kPa for 16 hours.
powder; odourless.
Assay. Weigh accurately about 0.75 g of the substance under
Identification examination, dissolve in 50 ml of anhydrous glacial acetic
acid. Titrate with 0.1 M perchloric acid, using crystal violet
A. Determine by infrared absorption spectrophotometry (2.4.6).
solution as indicator. Carry out a blank titration.
Compare the spectrum with that obtained with pentazocine
lactate RS or with the reference spectrum of pentazocine 1 ml of 0.1 M perchloric acid is equivalent to 0.03754 g of
lactate. C19H27NO,C3H6O3.
B. To 1 mg in a porcelain crucible add 0.5 ml of a solution of Storage. Store protected from light and moisture.
sulphuric acid containing 1 per cent w/v solution of
ammonium molybdate; an intense blue colour is produced
which changes to bluish green, green and finally, on standing,
yellow.
Pentazocine Injection
C. Gives reaction A of lactates (2.3.1).
Pentazocine Lactate Injection
Tests Pentazocine Injection is a sterile solution in Water for
pH (2.4.24). 5.5 to 6.5, determined in a 1.0 per cent w/v solution. Injections of either Pentazocine Lactate or pentazocine lactate
prepared by the interaction of Pentazocine and Lactic Acid.
Light absorption (2.4.7). Dissolve 0.1 g in 10 ml of 1 M
hydrochloric acid and add sufficient water to produce Pentazocine Injection contains not less than 95.0 per cent and
100 ml; dilute 10 ml to 100 ml with water. Absorbance of the not more than 105.0 per cent of the stated amount of
resulting solution at the maximum at about 278 nm, 0.50 to pentazocine, C19H27NO.
0.54. Description. A clear, colourless or almost colourless liquid.
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel HF254. Identification
Mobile phase. A mixture of 94 volumes of chloroform, A. To a volume containing 90 mg of pentazocine add 5 ml of
3 volumes of 2-propylamine and 3 volumes of methanol. 0.1 M sodium hydroxide and shake the resulting solution
with 5 ml of chloroform. Wash the chloroform extract with 2 ml
Test solution. Dissolve 0.2 g of the substance under
of water, dry over anhydrous sodium sulphate and filter.
examination in 10 ml of chloroform.
Evaporate the chloroform without applying heat and dry the
Reference solution (a). A 0.02 per cent w/v solution of the oily residue at a temperature not exceeding 25º and at a pressure
substance under examination in chloroform. of 2 kPa for 1 hour.

915
PENTOBARBITONE SODIUM IP 2007

On the residue, determine by infrared absorption 100.0 ml. To 5.0 ml add 1 ml of 1 M hydrochloric acid and
spectrophotometry (2.4.6). Compare the spectrum with that sufficient water to produce 100.0 ml. Measure the absorbance
obtained with pentazocine RS or with the reference spectrum of the resulting solution at the maximum at about 278 nm (2.4.7).
of pentazocine (form B). Calculate the content of C19H27NO, taking 69 as the specific
absorbance at 278 nm.
B. Dilute a volume containing 30 mg of pentazocine to 100 ml
with water. To 10 ml add 10 ml of 1 M sodium hydroxide and Storage. Store protected from light.
sufficient water to produce 100 ml. When examined in the
Labelling. The label states the strength in terms of the
range 230 nm to 360 nm (2.4.7), the solution shows absorption
equivalent amount of pentazocine in a suitable dose-volume.
maxima, at about 238 nm and 298 nm.
C. To a volume containing 30 mg of pentazocine add 2 ml of
0.1 M sodium hydroxide, extract with 2 ml of chloroform and
evaporate 0.1 ml of the chloroform extract to dryness in a
porcelain crucible. Add to the residue 0.5 ml of a 1 per cent
Pentobarbitone Sodium
w/v solution of ammonium molybdate in sulphuric acid; an Soluble Pentobarbitone; Pentobarbital Sodium
intense blue colour is produced which changes to bluish green,
green and finally, on standing, yellow.
H
O N ONa
Tests
H3C
pH (2.4.24). 4.0 to 5.0. N

Related substances. Determine by thin-layer chromatography H3C H3C O


(2.4.17), coating the plate with silica gel HF254.
Mobile phase. A mixture of 94 volumes of chloroform, C11H17N2NaO3 Mol. Wt. 248.3
3 volumes of 2-propylamine and 3 volumes of methanol.
Pentobarbitone Sodium is sodium 5-ethyl-5-[(1RS)-1-
Test solution. Dilute a volume of the injection with sufficient methylbutyl]barbiturate.
ethanol (95 per cent) to produce a solution containing
2.0 per cent w/v solution of pentazocine. Pentobarbitone Sodium contains not less than 99.0 per cent
and not more than 101.5 per cent of C11H17N2NaO3, calculated
Reference solution (a). Dilute 1 volume of the test solution to on the dried basis.
100 volumes with ethanol (95 per cent).
Description. A white, crystalline powder or granules;
Reference solution (b). Dilute 1 volume of the test solution to hygroscopic.
200 volumes with ethanol (95 per cent).
Reference solution (c). Dilute 1 volume of the test solution to Identification
400 volumes with ethanol (95 per cent). A. To 10 ml of a 10 per cent w/v solution add 5 ml of 2 M acetic
Apply to the plate 10 µl of each solution. After development, acid; a white, crystalline precipitate is produced. Filter, wash
dry the plate in air and examine in ultraviolet light at 254 nm. the precipitate with water and dry at 105º. Determine the
Heat the plate at 105º for 15 minutes, allow to cool, expose to melting point of the dried precipitate (2.4.21). Mix equal parts
iodine vapour and re-examine in ultraviolet light at 254 nm. of the dried precipitate and phenobarbitone RS and determine
Any secondary spot in the chromatogram obtained with the the melting point (2.4.21). The difference between the melting
test solution is not more intense than the spot in the points (which are about 131º) is not greater than 2º.
chromatogram obtained with reference solution (a), not more B. Complies with the test for identification of barbiturates
than one such spot is more intense than the spot in the (2.3.1), using the dried precipitate obtained in test A for
chromatogram obtained with reference solution (b) and not preparing the test solution.
more than four such spots are more intense than the spot in
the chromatogram obtained with reference solution (c). C. To 10 mg add 10 mg of vanillin and 2 ml of sulphuric acid,
mix and heat on a water-bath for 2 minutes; a reddish-brown
Other tests. Complies with the tests stated under Parenteral colour is produced. Cool and add 5 ml of ethanol; the colour
Preparations (Injections). changes to violet and then blue.
Assay. To an accurately measured volume containing about D. Ignite 1 g; the residue gives reaction A of sodium salts
0.15 g of pentazocine add sufficient water to produce (2.3.1).

916
IP 2007 PENTOBARBITONE TABLETS

Tests Pentobarbitone Tablets


Appearance of solution. Prepare freshly a 10.0 per cent w/v Pentobarbitone Sodium Tablets; Pentobarbital Sodium
solution in carbon dioxide-free water (solution A). Solution Tablets
A is clear (2.4.1).
Pentobarbitone Tablets contain not less than 92.5 per cent
pH (2.4.24). 9.6 to 11.0, determined in solution A. and not more than 107.5 per cent of the stated amount of
Related substances. Complies with the test for related pentobarbitone sodium, C11H17N2NaO3.
substances in barbiturates (2.3.4), but applying 10 µl of each
of the following solutions. Identification
Test solution. A 2.0 per cent w/v solution of the substance A. Shake a quantity of the powdered tablets containing 0.1 g
under examination. of Pentobarbitone Sodium with 10 ml of a 10 per cent w/v
solution of pyridine and filter. Add to the filtrate 1 ml of cupric
Reference solution. A 0.01 per cent w/v solution of the
sulphate with pyridine solution and set aside for 10 minutes;
substance under examination.
a reddish violet precipitate is produced.
Do not ignore any spot remaining on the line of application.
B. Shake a quantity of the powdered tablets containing 0.1 g
Free pentobarbitone. Not more than 3.5 per cent, determined of Pentobarbitone Sodium with 10 ml of water and filter. To
by the following method. Weigh accurately about 2.0 g and the filtrate add 2 ml of hydrochloric acid; a white precipitate
dissolve in 75 ml of dimethylformamide, heating gently if is produced (distinction from pentobarbitone).
necessary. Add 0.25 ml of a 1 per cent w/v solution of thymol C. The residue obtained in the Assay melts at 127º to
blue in dimethylformamide and titrate with 0.1 M sodium 130º (2.4.21).
methoxide to a blue end-point. Carry out a blank titration.
D. The powdered tablets, when moistened with hydrochloric
1 ml of 0.1 M sodium methoxide is equivalent to 0.02263 g of acid and introduced on a platinum wire into a flame, impart a
pentobarbitone. yellow colour to the flame.
Isomer. Dissolve 0.3 g in 5 ml of a 5 per cent w/v solution of
anhydrous sodium carbonate and add 0.3 g of 4-nitrobenzyl Tests
bromide dissolved in 10 ml of ethanol (95 per cent). Heat Isomer. Dissolve a quantity of the powdered tablets containing
under a reflux condenser for 30 minutes, cool to 25º scratch 0.3 g of Pentobarbitone Sodium in 5 ml of a 5 per cent w/v
the side of the vessel with a glass rod if necessary to induce solution of anhydrous sodium carbonate and add 0.3 g of
crystallisation and filter. Wash the residue with five quantities, 4-nitrobenzyl bromide dissolved in 10 ml of ethanol (95 per
each of 5 ml, of water. Transfer the residue as completely as cent). Heat under a reflux condenser for 30 minutes, cool to
possible to a small flask, add 25 ml of ethanol (95 per cent) 25º scratch the side of the vessel with a glass rod if necessary
and heat under a reflux condenser for 10 minutes; the solid to induce crystallisation and filter. Wash the residue with five
dissolves completely. Cool to 25º and scratch the side of the quantities, each of 5 ml, of water. Transfer the residue as
flask with a glass rod to induce crystallisation. Filter, wash the completely as possible to a small flask, add 25 ml of ethanol
residue with two quantities, each of 5 ml, of water and dry at (95 per cent) and heat under a reflux condenser for 10 minutes;
105º for 30 minutes. The dried residue melts at 136º to 148º filter the hot solution. Cool to 25º and scratch the side of the
(2.4.21). flask with a glass rod to induce crystallisation. Filter, wash the
Heavy metals (2.3.13). 0.67 g complies with the limit test for residue with two quantities, each of 5 ml, of water and dry at
heavy metals, Method B (30 ppm). 105º for 30 minutes. The dried residue melts at 136º to 148º
(2.4.21).
Loss on drying (2.4.19). Not more than 3.0 per cent, determined
on 1.0 g by drying in an oven at 105º. Other tests. Complies with the tests stated under Tablets.
Assay. Weigh accurately about 0.4 g, dissolve in 25 ml of a Assay. Weigh and powder 20 tablets. Weigh accurately a
12.75 per cent w/v solution of silver nitrate in pyridine and quantity of the powder containing about 0.3 g of
titrate with 0.1 M ethanolic sodium hydroxide using 0.5 ml of Pentobarbitone Sodium, dissolve as completely as possible
thymolphthalein solution as indicator, until a pure blue colour in 10 ml of a 2 per cent w/v solution of sodium hydroxide,
is obtained. Carry out a blank titration. saturate with sodium chloride, acidify with hydrochloric acid
and extract with successive quantities, each of 15 ml, of ether
1 ml of 0.1 M ethanolic sodium hydroxide is equivalent to until complete extraction is effected. Wash the combined
0.02483 g of C11H17N2NaO3. extracts with two quantities, each of 2 ml, of water and extract
Storage. Store protected from moisture. the combined washings with 10 ml of ether. Add the ether to

917
PEPSIN IP 2007

the main ether layer, filter and wash the filter with ether. 65 ml of 1 M hydrochloric acid to 1000 ml with water, continue
Evaporate the solvent and dry the residue to constant weight the trituration, dilute to 1000.0 ml with the acidified water and
at 105º. shake for 15 minutes. Prepare coagulated egg albumin by
1 g of residue is equivalent to 1.097 g of C11H17N2NaO3. boiling fresh hen-eggs in water for 15 minutes, cooling rapidly
to room temperature by immersion in cold water, separating
Storage. Store protected from moisture. the whites and rubbing through a no. 44 sieve. Reject the first
portion that passes through the sieve and triturate 15.0 g of
freshly prepared coagulated egg albumin with 50 ml of the
Pepsin acidified water ensuring that the particles of egg albumin are
thoroughly disintegrated, add a further 50 ml of the acidified
Pepsin is obtained from the gastric mucosa of pigs, cattle or water and keep in a water-bath at 51º ± 1º for 15 minutes. Add
sheep. It contains gastric proteinases that are active in an 20.0 ml of the prepared solution of the substance under
acid medium, pH 1 to 5. It may contain a suitable diluent such examination and digest at 51º ± 1º for 4 hours, shaking at
as Lactose. intervals of 15 minutes. Centrifuge and decant off most of the
Pepsin has an activity equivalent to its ability to digest not clear supernatant liquid, wash the remainder into a 10-ml
less than 3000 times its weight of coagulated egg albumin graduated cylinder and allow to stand for 30 minutes. The
when determined by the method given under Assay. volume of the undissolved albumin is not more than 2 ml.
Description. A white or light buff-coloured, crystalline or Storage. Store protected from moisture.
amorphous powder or translucent scales; odour, faint and
meaty but not rancid; hygroscopic.

Identification Peritoneal Dialysis Solutions


A. Place 1 ml of congo red fibrin on a filter paper and wash Intraperitoneal Dialysis Fluids
until a colourless filtrate is obtained with a solution prepared
by diluting 30 ml of 1 M hydrochloric acid to 1000 ml with Peritoneal Dialysis Solutions are sterile preparations for
water and adjusting the pH 1.5 to 1.7. Perforate the filter paper intraperitoneal use containing electrolytes with a concentration
and wash the congo red fibrin through it with 20 ml of the close to the electrolytic composition of plasma. They contain
same hydrochloric acid solution. Shake this suspension before dextrose in varying concentrations and/or other suitable
use. Dissolve about 10 mg of the substance under examination osmotic agents. They do not contain antioxidants such as
in 2 ml of the hydrochloric acid solution and adjust the pH 1.5 metabisulphite salts.
to 1.7. Place 4 ml of the congo red fibrin suspension in each of Peritoneal Dialysis Solutions contain not less than 97.5 per
two tubes. To one of the tubes add 1 ml of the solution of the cent and not more than 102.5 per cent of the stated amount of
substance under examination and to the other tube add 1 ml of sodium, Na, not less than 95.0 per cent and not more than
water (control solution). Mix the contents of each tube and 105.0 per cent of the stated amounts of potassium, K, calcium,
place in a water-bath at 25º with gentle shaking for 15 minutes; Ca, magnesium, Mg, chloride, Cl, acetate, C2H3O2, lactate,
the control solution is colourless and the solution of the C3H5O3, sodium bicarbonate, NaHCO3, bicarbonate, HCO3 and
substance under examination is violet blue. dextrose, C6H12O6.
B. The proteolytic activity of a solution in water is destroyed Description. Clear, colourless or faintly straw-coloured
at once by boiling. It is destroyed by warming for 10 minutes solutions.
at 40º at a pH of 8.0.
Identification
Tests
A. To 5 ml of the solution under examination, add 2 ml of
Microbial contamination (2.2.9). 1 g is free from Escherichia
dilute sodium hydroxide solution and 0.05 ml of potassium
coli and 10 g is free from salmonellae.
cupri-tartrate solution; the solution remains blue and clear.
Sulphated ash (2.3.18). Not more than 5.0 per cent. Heat to boiling; a copious red precipitate is formed.
Loss on drying (2.4.19). Not more than 5.0 per cent, determined B. 20 ml gives reactions of chlorides, sodium salts, potassium
on 1.0 g by drying in an oven at 60º at a pressure not exceeding salts and calcium salts (2.3.1).
0.7 kPa for 4 hours.
C. To 5 ml add 1 ml of hydrochloric acid in a test-tube fitted
Assay. Weigh accurately 0.25 g, triturate with 1.0 g of sodium with a stopper and a bent tube, heat and collect a few ml of the
chloride, add slowly acidified water prepared by diluting distillate. The distillate gives reaction C of acetates (2.3.1).

918
IP 2007 PERITONEAL DIALYSIS SOLUTIONS

D. To 0.1 ml of titan yellow solution add 10 ml of water, 2 ml of Pyrogens (2.2.8). Solutions for which a validated test for
the solution under examination and 1 ml of 1 M sodium bacterial endotoxins cannot be carried out, comply with the
hydroxide; a pink colour is produced if magnesium salts are test for pyrogens, injecting 10 ml of the solution per kg of the
present. rabbit’s body weight.
E. Lactates and bicarbonates are identified together with the Sterility (2.2.11). Complies with the test for sterility.
Assay for lactate and bicarbonate. Assay. For sodium — Dilute suitably with water and determine
by Method A for flame photometry (2.4.4), or by Method A for
Tests atomic absorption spectrophotometry (2.4.2), measuring at
589 nm and using sodium solution FP, or sodium solution
Appearance of solution. The solution under examination is
AAS respectively, suitably diluted with water for the standard
clear (2.4.1), and not more intensely coloured than reference
solutions.
solution YS4 (2.4.1).
For potassium — Dilute suitably with water and determine
pH (2.4.24). 4.5 to 6.5. If the solution contains bicarbonate, 6.5 by Method A for flame photometry (2.4.4), or by Method A for
to 8.0. atomic absorption spectrophotometry (2.4.2), measuring at
5-Hydroxymethylfurfural and Related substances. Dilute a 767 nm and using potassium solution FP or potassium solution
volume containing 1.0 g of Dextrose to 250.0 ml with water AAS respectively, suitably diluted with water for the standard
and measure the absorbance of the resulting solution (2.4.7) solutions.
at the maximum at about 284 nm; absorbance at about 284 nm, For calcium — Dilute suitably with water and determine by
not more than 0.25. Method A for flame photometry (2.4.4), or by Method A for
Aluminium. Adjust the pH of 400 ml of the solution under atomic absorption spectrophotometry (2.4.2), measuring at
examination to pH 6.0 and add 10 ml of acetate buffer pH 6.0. 422.7 nm and using calcium solution FP or calcium solution
Extract the resulting solution with successive quantities of AAS respectively, suitably diluted with water for the standard
20, 20 and 10 ml of a 0.5 per cent w/v solution of solutions.
8-hydroxyquinoline in chloroform and dilute the combined For magnesium — To 50.0 ml add 50 ml of water and 5 ml of
extracts to 50.0 ml with chloroform. Use as the blank a mixture strong ammonia-ammonium chloride solution and titrate with
of 10 ml of acetate buffer pH 6.0 and 100 ml of water treated in 0.005 M disodium edetate using 50 mg of eriochrome black
the same manner and as the standard solution a mixture of T mixture as indicator.
2.0 ml of aluminium standard solution (2 ppm Al), 10 ml of 1 ml of 0.005 M disodium edetate is equivalent to 0.1215 mg
acetate buffer pH 6.0 and 90 ml of water treated in the same of Mg.
manner. Measure the fluorescence of the test solution (I1), of
For total chloride — Dilute an accurately measured volume
the standard solution (I2) and of the blank (I3), (2.4.5), using an
containing about 60 mg of chloride to 50.0 ml with water. Add
excitation wavelength of 392 nm and a secondary filter with a
25.0 ml of 0.1 M silver nitrate and 2 ml of nitric acid. Filter,
transmission band centred at 518 nm, or a monochromator set
wash the precipitate with water slightly acidified with nitric
to transmit at this wavelength. The fluorescence of the test
acid and titrate the excess of silver nitrate with 0.1 M
solution (I1 – I3) is not greater than that of the standard solution
ammonium thiocyanate using ferric ammonium sulphate
(I2 – I3).
solution as indicator until a reddish yellow colour is produced.
Particulate contamination (2.5.9). Carry out the test using Carry out a blank titration.
50 ml of the solution under examination. 1 ml of 0.1 M silver nitrate is equivalent to 0.003545 g of total
The preparation meets the requirements of the test if it contains chloride, calculated as Cl.
particles within the maximum limits shown below. For acetate (if present) — Determine by liquid
chromatography (2.4.14).
Particle size in µm Maximum number
(Equal to or larger than) of particles per ml Test solution. Dilute an accurately measured volume of the
preparation under examination quantitatively with water to
10 25 obtain a solution containing about 1.0 mg of acetate per ml.
25 3 Reference solution. Dissolve an accurately weighed quantity
of sodium acetate in water to obtain a solution having a known
Other tests. Complies with the tests stated under Parenteral concentration of about 0.12 per cent w/v of Sodium Acetate.
Preparations (Injections).
Chromatographic system
Bacterial endotoxins (2.2.3). Not more than 0.25 Endotoxin – a stainless steel column 30 cm x 7.8 mm, packed with a
Unit per ml. strong cation-exchange resin consisting of sulphonated

919
PERITONEAL DIALYSIS SOLUTIONS IP 2007

cross-linked styrene-divinylbenzene copolymer in the 1 ml of 0.1 M hydrochloric acid is equivalent to 8.40 mg of


hydrogen form (7 µm), NaHCO3.
– mobile phase: filtered and degassed 0.1 M sulphuric For lactate and bicarbonate — Determine by liquid
acid, chromatography (2.4.14).
– flow rate. 0.8 ml per minute,
Test solution. Use the preparation under examination.
– column temperature. 60º,
– spectrophotometer set at 210 nm, Reference solutions. Dissolve accurately weighed quantities
– a 20 µl loop injector. of lactates and bicarbonates in order to obtain solutions having
concentrations of about 90 per cent, 100 per cent and 110 per
Inject the reference solution and record the chromatograms. cent of the claim in 100 ml of water.
The test is not valid unless the tailing factor is not more than
2.0 and the relative standard deviation for replicate injections Chromatographic system
is not more than 2.0 per cent. – a stainless steel column 30 cm x 7.8 mm, packed with a
cation-exchange resin (9 µm),
Inject separately the test and standard solutions and record – column. temperature 85º,
the chromatograms. Measure the responses for the major peak – mobile phase: filtered and degassed 0.005 M sulphuric
and calculate the content of acetate in the preparation under acid,
examination. – flow rate. 0.6 ml per minute,
For lactate (if present) — Determine by liquid chromatography – differential refractometer detector,
(2.4.14). – a 20 µl loop injector.
Test solution. Use the preparation under examination. Inject separately the test solution and the reference solutions
in duplicate, and record the chromatograms in the prescribed
Reference solution(a). Dissolve an accurately weighed conditions. The peaks elute in the following order, lactates,
quantity of sodium lactate RS in water to obtain a solution then bicarbonates. Determine the concentration of lactates
having a known concentration of about 2 mg per ml. and bicarbonates in the test solution by interpolating the peak
Reference solution (b). Prepare a solution in water containing area for lactate and the peak height for bicarbonate from the
about 3 mg of anhydrous sodium acetate and 3 mg of sodium linear regression curve obtained with the solutions prepared
lactate RS per ml. as reference solutions.
Chromatographic system For dextrose — Transfer a volume of the preparation under
– a stainless steel column 10 cm x 4.6 mm, packed with examination containing about 25 mg of Dextrose to a 250-ml
octadecylsilane chemically bonded to porous silica or conical flask with a ground-glass neck and add 25.0 ml of
ceramic microparticles 3 to 10 µm, cupri-citric solution. Add a few grains of pumice, fit a reflux
– mobile phase: a filtered and degassed solution in water condenser, heat so that boiling occurs within 2 minutes and
containing about 1 ml of formic acid and 1 ml of boil for exactly 10 minutes. Cool and add 3 g of potassium
dicyclohexylamine per litre, iodide dissolved in 3 ml of water. Carefully add, in small
– flow rate. 1 ml per minute, amounts, 25 ml of a 25 per cent w/w solution of sulphuric acid.
– spectrophotometer set at 210 nm, Titrate with 0.1 M sodium thiosulphate using starch solution
– a 20 µl loop injector. as indicator. Carry out a blank titration using 25 ml of water.
Calculate the content of anhydrous dextrose, C6H12O6, from
Inject separately reference solutions (a) and (b), and record
the following Table.
the chromatograms. The test is not valid unless the resolution
between the peaks due to acetate and lactate is not less than Volume of Anhydrous dextrose
2.0, the tailing factor for the analyte peak is not more than 2.0 0.1 M sodium thiosulphate
and the relative standard deviation for replicate injections is consumed (ml) (mg)
not more than 2.0 per cent. 8 19.8
Inject separately the test solution and reference solution (a), 9 22.4
and record the chromatograms. Measure the responses for 10 25.0
the major peak and calculate the content of lactate in the 11 27.6
preparation under examination. 12 30.3
For sodium bicarbonate — Titrate with 0.1 M hydrochloric 13 33.0
acid a volume of the preparation under examination containing 14 35.7
about 0.1 g of sodium bicarbonate, determining the end-point 15 38.5
potentiometrically (2.4.25). 16 41.3

920
IP 2007 PETHIDINE HYDROCHLORIDE

Storage. Peritoneal Dialysis Solutions are supplied in rigid or B. To 5 ml of a 1 per cent w/v solution add a few drops of
semi-rigid plastic containers, in flexible plastic containers fitted potassium mercuri-iodide solution; a cream-coloured
with a special connecting device (these are generally filled to precipitate is produced.
a volume below their nominal capacity and presented in closed
C. Dissolve 5 mg in 0.5 ml of water and add 0.1 ml of
protective envelopes) or in glass containers. Store at a
formaldehyde solution and 2 ml of sulphuric acid; an orange-
temperature not exceeding 30º.
red colour is produced.
CAUTION — Exposure to temperatures below 4º may cause
D. Gives reaction A of chlorides (2.3.1).
crystallisation and separation of solid particles rendering
the preparation unsuitable for use.
Tests
Labelling. The label states (1) the formula of the solution for
peritoneal dialysis, expressed in grams per litre and in millimoles Appearance of solution. A 2.0 per cent w/v solution in carbon
per litre; (2) the total osmolar concentration in mOsmol per dioxide-free water is clear (2.4.1), and colourless (2.4.1).
litre; (3) the nominal volume of the solution in the container; Acidity or alkalinity. To 10 ml of a 2.0 per cent w/v solution in
(4) that the solution is free from bacterial endotoxins, or where carbon dioxide-free water add 0.2 ml of methyl red solution
applicable, that it is apyrogenic; (5) that the solution is not to and 0.2 ml of 0.01 M sodium hydroxide; the solution is yellow.
be used for intravenous infusion; (6) that any unused portion Add 0.3 ml of 0.01 M hydrochloric acid; the solution is red.
of the solution is to be discarded; (7) that the solution
containing visible particles should not be used; (8) the storage Related substances. Determine by thin-layer chromatography
conditions. (2.4.17), coating the plate with kieselguhr G.
Mobile phase. The upper layer obtained by shaking together
100 volumes of light petroleum (50º to 70º), 8 volumes of
2-phenoxyethanol and 1 volume of diethylamine.
Pethidine Hydrochloride
Test solution. Dissolve 0.1 g in 5 ml of water, add 0.5 ml of
Meperidine Hydrochloride 10 M sodium hydroxide and 2 ml of ether and shake; allow the
layers to separate and use the upper layer.
CH3 Reference solution. Dilute 0.5 ml of the test solution to 50 ml
N with ether.
Impregnate the dry plate by placing it in a closed tank
O CH3 , HCl containing a mixture of 90 volumes of acetone and 10 volumes
of 2-phenoxyethanol so that the plate dips about 5 mm beneath
O
the surface of the liquid, allowing the impregnating solvent to
ascend at least 15 cm, removing the plate from the tank and
drying in a current of air. Use immediately, with the flow of the
C15H21NO2,HCl Mol. Wt. 283.8 mobile phase in the same direction as the impregnation.
Pethidine Hydrochloride is ethyl 1-methyl-4- Apply to the plate 5 µl of each solution. Allow the mobile
phenylpiperidine-4-carboxylate hydrochloride. phase to rise 12 cm. Dry the plate in air for 10 minutes, return
the plate to the tank and repeat the development. Remove the
Pethidine Hydrochloride contains not less than 99.0 per cent
plate, allow it to dry in air for 10 minutes and spray with a
and not more than 101.0 per cent of C15H21NO2,HCl, calculated
0.2 per cent w/v solution of 2,7-dichlorofluorescein in
on the dried basis.
methanol. Allow to stand for 5 minutes and spray with water
Description. Colourless crystals or a white, crystalline powder. until the background is white to pale yellow. Examine in
daylight. The chromatograms show red or orange spots. Any
Identification secondary spot in the chromatogram obtained with the test
solution is not more intense than the spot in the chromatogram
Test A may be omitted if tests B, C and D are carried out. Tests
obtained with the reference solution. Examine without delay
B and C may be omitted if tests A and D are carried out.
in ultraviolet light at 365 nm. The chromatograms show spots
A. Determine by infrared absorption spectrophotometry (2.4.6). with intense yellow fluorescence. Any secondary spot in the
Compare the spectrum with that obtained with pethidine chromatogram obtained with the test solution is not more
hydrochloride RS or with the reference spectrum of pethidine intense than the spot in the chromatogram obtained with the
hydrochloride. reference solution.

921
PETHIDINE INJECTIONS IP 2007

Sulphated ash (2.3.18). Not more than 0.1 per cent. necessary to 5 ml with water, with 0.5 ml of 5 M sodium
Loss on drying (2.4.19). Not more than 0.5 per cent, determined hydroxide and 2 ml of ether.
on 1.0 g by drying in an oven at 105º. Reference solution. Dilute 0.5 ml of the test solution to 50 ml
with ether.
Assay. Weigh accurately about 0.5 g, dissolve in 30 ml of
anhydrous glacial acetic acid, add 12 ml of mercuric acetate Impregnate the dry plate by placing it in a closed tank
solution. Titrate with 0.1 M perchloric acid, using crystal containing a mixture of 90 volumes of acetone and 10 volumes
violet solution as indicator. Carry out a blank titration. of 2-phenoxyethanol so that the plate dips about 5 mm beneath
the surface of the liquid, allowing the impregnating solvent to
1 ml of 0.1 M perchloric acid is equivalent to 0.02838 g of
ascend at least 15 cm, removing the plate from the tank and
C15H21NO2,HCl.
drying in a current of air. Use immediately, with the flow of the
Storage. Store protected from light and moisture. mobile phase in the same direction as the impregnation.
Apply to the plate 5 µl of each solution. Allow the mobile
phase to rise 12 cm. Dry the plate in air for 10 minutes, return
Pethidine Injection the plate to the tank and repeat the development. Remove the
plate, allow it to dry in air for 10 minutes and spray with a
Pethidine Hydrochloride Injection; Meperidine 0.2 per cent w/v solution of 2,7-dichlorofluorescein in
Hydrochloride Injection methanol. Allow to stand for 5 minutes and spray with water
Pethidine Injection is a sterile solution of Pethidine until the background is white to pale yellow. Examine in
Hydrochloride in Water for Injections. daylight. The chromatograms show red or orange spots. Any
secondary spot in the chromatogram obtained with the test
Pethidine Injection contains not less than 95.0 per cent and solution is not more intense than the spot in the chromatogram
not more than 105.0 per cent of pethidine hydrochloride, obtained with the reference solution. Examine without delay
C15H21NO2,HCl. in ultraviolet light at 365 nm. The chromatograms show spots
with intense yellow fluorescence. Any secondary spot in the
Identification
chromatogram obtained with the test solution is not more
A. To a volume containing 50 mg of Pethidine Hydrochloride intense than the spot in the chromatogram obtained with the
add sufficient 1 M sodium hydroxide to make strongly alkaline reference solution.
to litmus paper and extract with two quantities, each of 10 ml, Other tests. Complies with the tests stated under Parenteral
of chloroform. Wash the combined extracts with 5 ml of water, Preparations (Injections).
dry over anhydrous sodium sulphate, filter and evaporate the
Assay. Dilute an accurately measured volume containing about
filtrate to dryness. Remove the last traces of chloroform by
0.1 g of Pethidine Hydrochloride with 40 ml of water, add 1 ml
drying the residual oil at 60º at a pressure not exceeding
of 5 M sodium hydroxide and extract immediately with
0.7 kPa.
successive quantities of 25, 10 and 10 ml of chloroform. Wash
On the oily residue, determine by infrared absorption each extract with the same 15 ml of water, filter into a dry flask
spectrophotometry (2.4.6). Compare the spectrum with that and combine the extracts (which should be clear and free from
obtained with pethidine hydrochloride RS or with the droplets of water). Titrate with 0.02 M perchloric acid, using
reference spectrum of pethidine. 0.15 ml of 1-naphtholbenzein solution as indicator. Carry out
B. To 0.5 ml add 0.1 ml of formaldehyde solution and 2 ml of a blank titration.
sulphuric acid; an orange-red colour is produced. 1 ml of 0.02 M perchloric acid is equivalent to 0.005676 g of
C. Gives the reactions of chlorides (2.3.1). C15H21NO2,HCl.
Storage. Store protected from light.
Tests
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with kieselguhr G. Pethidine Tablets
Mobile phase. The upper layer obtained by shaking together Pethidine Hydrochloride Tablets; Meperidine
100 volumes of light petroleum (50º to 70º), 8 volumes of Hydrochloride Tablets
2-phenoxyethanol and 1 volume of diethylamine. Pethidine Tablets contain not less than 92.5 per cent and not
Test solution. The upper layer obtained by shaking a volume more than 107.5 per cent of the stated amount of pethidine
containing 0.1 g of Pethidine Hydrochloride, diluted if hydrochloride, C15H21NO2,HCl.

922
IP 2007 PHENINDAMINE TARTRATE

Identification intense than the spot in the chromatogram obtained with the
reference solution.
A. Shake a quantity of the powdered tablets containing 50 mg
of Pethidine Hydrochloride with 20 ml of chloroform, filter, Other tests. Comply with the tests stated under Tablets.
evaporate the filtrate to dryness and dry the residue at a Assay. Weigh and powder 20 tablets. Weigh accurately a
pressure of 2 kPa. quantity of the powder containing about 0.3 g of Pethidine
On the residue, determine by infrared absorption Hydrochloride in 40 ml of water, add 2 ml of 5 M sodium
spectrophotometry (2.4.6). Compare the spectrum with that hydroxide and extract immediately with successive quantities
obtained with pethidine hydrochloride RS or with the of 25, 10 and 10 ml of chloroform. Wash each extract with the
reference spectrum of pethidine hydrochloride. same 15 ml of water and filter into a dry flask. Combine the
extracts (which should be clear and free from droplets of water).
B. Shake a quantity of the powdered tablets containing 0.2 g Titrate with 0.05 M perchloric acid, using 0.15 ml of
of Pethidine Hydrochloride with 20 ml of water and filter. To 1-naphtholbenzein solution as indicator. Carry out a blank
5 ml of the filtrate add 10 ml of picric acid solution. The crystals titration.
so obtained, after washing with water and drying, melt at
about 190º (2.4.21). 1 ml of 0.05 M perchloric acid is equivalent to 0.01419 g of
C15H21NO2,HCl.
Tests Storage. Store protected from light and moisture.
Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with kieselguhr G.
Mobile phase. The upper layer obtained by shaking together Phenindamine Tartrate
100 volumes of light petroleum (50º to 70º), 8 volumes of
2-phenoxyethanol and 1 volume of diethylamine.
Test solution. The upper layer obtained by shaking a quantity
of the powdered tablets containing 0.1 g of Pethidine
Hydrochloride with 5 ml of water, filtering, shaking the filtrate H OH
with 0.5 ml of 5 M sodium hydroxide and 2 ml of ether and , COOH
allowing the layers to separate. N CH3 HOOC
H OH
Reference solution. Dilute 0.5 ml of the test solution to 50 ml
with ether.
C19H19N,C4H6O6 Mol. Wt. 411.5
Impregnate the dry plate by placing it in a closed tank
Phenindamine Tartrate is (RS)-2,3,4,9-tetrahydro-2-methyl-9-
containing a mixture of 90 volumes of acetone and 10 volumes
phenyl-1H-indeno[2,1-c]pyridine (2R,3R)-tartrate.
of 2-phenoxyethanol so that the plate dips about 5 mm beneath
the surface of the liquid, allowing the impregnating solvent to Phenindamine Tartrate contains not less than 98.5 per cent
ascend at least 15 cm, removing the plate from the tank and and not more than 101.0 per cent of C19H19N, C4H6O6, calculated
drying in a current of air. Use immediately, with the flow of the on the dried basis.
mobile phase in the same direction as the impregnation. Description. A white or almost white voluminous powder;
Apply to the plate 5 µl of each solution. Allow the mobile odourless or almost odourless.
phase to rise 12 cm. Dry the plate in air for 10 minutes, return
the plate to the tank and repeat the development. Remove the Identification
plate, allow it to dry in air for 10 minutes and spray with a A. When examined in the range 230 nm to 360 nm (2.4.7), a
0.2 per cent w/v solution of 2,7-dichlorofluorescein in 0.004 per cent w/v solution shows an absorption maximum
methanol. Allow to stand for 5 minutes and spray with water only at about 259 nm; absorbance at about 259 nm, about 0.88.
until the background is white to pale yellow. Examine in
daylight. The chromatograms show red or orange spots. Any B. Dissolve 25 mg in 5 ml of sulphuric acid; an orange-brown
secondary spot in the chromatogram obtained with the test colour is produced which is discharged when the solution is
solution is not more intense than the spot in the chromatogram carefully diluted with 20 ml of water.
obtained with the reference solution. Examine without delay C. Dissolve 0.5 g in 15 ml of hot water, add a slight excess of
in ultraviolet light at 365 nm. The chromatograms show spots 5 M sodium hydroxide, filter and neutralise the filtrate to litmus
with intense yellow fluorescence. Any secondary spot in the paper with 2 M hydrochloric acid. The solution gives reaction
chromatogram obtained with the test solution is not more B of tartrates (2.3.1).

923
PHENINDAMINE TABLETS IP 2007

D. Melting range (2.4.21). 160º to 162º, when heated to 163º it accurately measured quantity of the filtrate containing about
re-solidifies and melts again at about 168º, with decomposition. 0.2 g of Phenindamine Tartrate add 10 ml of 5 M sodium
hydroxide, and extract with 25 ml and then with three quantities,
Tests each of 10 ml, of chloroform, washing each extract with the
same 15 ml of water and filtering in a dry flask. Combine the
pH (2.4.24). 3.4 to 3.9, determined in a 1.0 per cent w/v solution.
extracts (which should be clear and free from droplets of
Sulphated ash (2.3.18). Not more than 0.1 per cent. water).Titrate with 0.02 M perchloric acid, using oracet blue
Loss on drying (2.4.19). Not more than 1.0 per cent, determined B solution as indicator. Carry out a blank titration.
on 1.0 g by drying in an oven at 105º. 1 ml of 0.02 M perchloric acid is equivalent to 0.008229 g of
Assay. Weigh accurately about 0.4 g, dissolve in 40 ml of C19H19N, C4H6O6.
warm water, cool, add 10 ml of dilute sodium carbonate Storage. Store protected from light and moisture.
solution and extract with successive quantities of 25, 10 and
10 ml of chloroform, washing each extract with the same 15 ml
of water and filtering into a dry flask. Combine the extracts
(which should be clear and free from droplets of water). Titrate
Phenindione
with 0.05 M perchloric acid, using oracet blue B solution as O
indicator. Carry out a blank titration.
1 ml of 0.05 M perchloric acid is equivalent to 0.02057 g of
C19H19N, C4H6O6.
Storage. Store protected from light and moisture. O
C15H10O2 Mol. Wt. 222.2
Phenindione is 2-phenylindane-1,3-dione.
Phenindamine Tablets Phenindione contains not less than 98.0 per cent and not
Phenindamine Tartrate Tablets more than 100.5 per cent of C15H10O2, calculated on the dried
basis.
Phenindamine Tablets contain not less than 92.5 per cent and
Description. Soft, white or creamy-white crystals; almost
not more than 107.5 per cent of the stated amount of
odourless.
phenindamine tartrate, C19H19N, C4H6O6. The tablets are coated.
Identification
Identification
A. Determine by infrared absorption spectrophotometry (2.4.6).
A. Shake a quantity of the powdered tablets containing 20 mg Compare the spectrum with that obtained with phenindione
of Phenindamine Tartrate with 100 ml of water, dilute 10 ml to RS.
100 ml with water and filter; absorbance of the filtrate at the
maximum at about 259 nm, about 0.44 (2.4.7). B. Dissolve 0.1 g in 30 ml of ethanol (95 per cent) with the aid
of heat, cool and add sufficient ethanol (95 per cent) to
B. To a portion of the finely powdered tablets containing about produce 50 ml. Dilute 10 ml of this solution to 250 ml with
50 mg of Phenindamine Tartrate, add 5 ml of water and 3 ml of 0.1 M sodium hydroxide and further dilute 5 ml to 100 ml with
dilute ammonia solution and extract the liberated base with 0.1 M sodium hydroxide. When examined in the range 230 nm
two successive quantities, each of 10 ml, of ether. Wash the and 360 nm (2.4.7), the solution shows absorption maxima at
combined ether extracts with 2 ml of water and evaporate to about 278 nm and at about 330 nm; absorbance at about
dryness. Dissolve 25 mg of the residue in 5 ml of sulphuric 278 nm, about 0.55 and at about 330 nm, about 0.16.
acid; an orange-brown colour is produced which is discharged
when the solution is carefully diluted with 20 ml of water. C. To 1 g add 50 ml of ethanol (95 per cent) and 0.5 ml of
aniline, heat gently under a reflux condenser for 3 hours, cool
Tests in ice and filter. The residue, after washing with 2 ml of ethanol
(95 per cent) and recrystallisation from chloroform, melts at
Other tests. Complies with the tests stated under Tablets. about 225º (2.4.21).
Assay. Weigh and digest 20 tablets with 70 ml of water and
Tests
5 ml of dilute hydrochloric acid until completely disintegrated,
filter and wash the residue with 20 ml of water. Dilute the Related substances. Determine by thin-layer chromatography
combined filtrate and washings to 100.0 ml with water. To an (2.4.17), coating the plate with silica gel GF254.

924
IP 2007 PHENINDIONE TABLETS

Mobile phase. A 0.02 per cent w/v solution of butylated A. Determine by infrared absorption spectrophotometry (2.4.6).
hydroxytoluene in a mixture of 80 volumes of toluene, Compare the spectrum with that obtained with phenindione
20 volumes of ethyl acetate and 4 volumes of glacial acetic RS.
acid. B. Dissolve 0.1 g in 30 ml of ethanol (95 per cent) with the aid
Test solution. Dissolve 0.1 g of the substance under of heat, cool and add sufficient ethanol (95 per cent) to
examination in 10 ml of dichloromethane. produce 50 ml. Dilute 10 ml of this solution to 250 ml with
0.1 M sodium hydroxide and further dilute 5 ml to 100 ml with
Reference solution (a). A 0.02 per cent w/v solution of the 0.1 M sodium hydroxide. When examined in the range 230 nm
substance under examination in dichloromethane. to 360 nm (2.4.7), the solution shows absorption maxima at
Reference solution (b). A 0.005 per cent w/v solution of the about 278 nm and 330 nm; absorbance at about 278. about
substance under examination in dichloromethane. 0.55 and at about 330 nm, about 0.16.

Apply to the plate 10 µl of each solution. Allow the mobile C. To 50 mg add 1 ml of sulphuric acid; a deep blue to violet
phase to rise 4 cm. Dry the plate in warm air and examine in solution is produced. On dilution with water the solution
ultraviolet light at 254 nm. Any secondary spot in the becomes colourless and a white precipitate is produced.
chromatogram obtained with the test solution is not more
Tests
intense than the spot in the chromatogram obtained with
reference solution (a) and not more than one such spot is Related substances. Determine by thin-layer chromatography
more intense than the spot in the chromatogram obtained (2.4.17), coating the plate with silica gel GF254.
with reference solution (b).
Mobile phase. A 0.02 per cent w/v solution of butylated
Sulphated ash (2.3.18). Not more than 0.1 per cent. hydroxytoluene in a mixture of 80 volumes of toluene,
20 volumes of ethyl acetate and 4 volumes of glacial acetic
Loss on drying (2.4.19). Not more than 1.0 per cent, determined
acid.
on 1.0 g by drying in oven at 105º for 2 hours.
Test solution. Shake a quantity of the powdered tablets
Assay. Weigh accurately about 0.3 g, add 50 ml of ethanol
containing 50 mg of Phenindione with 15 ml of
(95 per cent) and warm until solution is effected. Cool to
dichloromethane, filter, evaporate the filtrate to dryness and
room temperature, add 10 ml of a 10 per cent v/v solution of
dissolve the residue in 5 ml of dichloromethane.
bromine in ethanol (95 per cent) and allow to stand for
10 minutes, shaking occasionally. Add 1 g of 2-naphthol and Reference solution (a). Dilute 1 volume of the test solution to
shake until the colour of the bromine is discharged. Remove 50 volumes with dichloromethane.
any vapour of bromine in the flask with a current of air, add Reference solution (b). Dilute 1 volume of the test solution to
50 ml of water and 10 ml of dilute potassium iodide solution 200 volumes with dichloromethane.
and titrate the liberated iodine with 0.1 M sodium thiosulphate
using starch solution as indicator. Apply to the plate 10 µl of each solution. Allow the mobile
phase to rise 4 cm. Dry the plate in warm air and examine in
1 ml of 0.1 M sodium thiosulphate is equivalent to 0.01111 g ultraviolet light at 254 nm. Any secondary spot in the
of C15H10O2. chromatogram obtained with the test solution is not more
Storage. Store protected from moisture. intense than the spot in the chromatogram obtained with
reference solution (a) and not more than one such spot is
more intense than the spot in the chromatogram obtained
with reference solution (b).
Phenindione Tablets Uniformity of content. (For tablets containing 50 mg or less)
— Comply with the test stated under Tablets.
Phenindione Tablets contain not less than 92.5 per cent and
not more than 107.5 per cent of the stated amount of Place one tablet in 50 ml of 0.1 M sodium hydroxide, dissolve
phenindione, C15H10O2. completely by shaking gently, add a further 100 ml of 0.1 M
sodium hydroxide and shake for 1 hour. Dilute to 250.0 ml with
Identification 0.1 M sodium hydroxide, filter and dilute a portion of the
filtrate with sufficient 0.1 M sodium hydroxide to produce a
Shake a quantity of the powdered tablets containing 0.2 g of solution containing 4 µg of Phenindione per ml. Measure the
Phenindione with 50 ml of chloroform, filter and evaporate the absorbance of the resulting solution at the maximum at about
filtrate to dryness. Recrystallise the residue from ethanol 278 nm (2.4.7). Calculate the content of C15H10O2 taking 1310
(95 per cent). The crystals complies with the following tests. as the specific absorbance at 278 nm.

925
PHENIRAMINE MALEATE IP 2007

Other tests. Complies with the tests stated under Tablets. add 5 ml of water; a yellow colour is produced. To 2 ml of the
Assay. Weigh and powder 20 Tablets. Weigh accurately a solution add 3 ml of a 50 per cent w/v solution of ammonium
quantity of the powder containing about 50 mg of Phenindione acetate, previously cooled in ice; a pink colour is produced
and shake with 150 ml of 0.1 M sodium hydroxide for 1 hour, which persists for at least 10 minutes in the cooled solution.
add sufficient 0.1 M sodium hydroxide to produce 250.0 ml, Tests
filter and dilute 5.0 ml of the filtrate to 250.0 ml with 0.1 M
sodium hydroxide. Measure the absorbance of the resulting pH (2.4.24). 4.5 to 5.5, determined in a 1.0 per cent w/v solution.
solution at the maximum at about 278 nm (2.4.7). Calculate the
Related substances. Determine by thin-layer chromatography
content of C15H10O2 taking 1310 as the specific absorbance at
(2.4.17), coating the plate with silica gel G.
278 nm.
Mobile phase. A mixture of 50 volumes of cyclohexane,
Storage. Store protected from moisture.
40 volumes of chloroform and 10 volumes of diethylamine.
Test solution. Dissolve 0.2 g of the substance under
examination in 10 ml of methanol.
Pheniramine Maleate
Reference solution (a). A 0.02 per cent w/v solution of the
substance under examination in methanol.
Reference solution (b). A 0.004 per cent w/v solution of the
substance under examination in methanol.
COOH
N CH3 , Apply to the plate 10 µl of each solution. After development,
N
dry the plate in air, spray with dilute potassium
CH3 COOH
iodobismuthate solution. Any secondary spot in the
chromatogram obtained with the test solution is not more
C16H20N2,C4H4O4 Mol. Wt. 356.4 intense than the spot in the chromatogram obtained with
Pheniramine Maleate is (3RS)-N,N-dimethyl-3-phenyl-3- reference solution (a) and not more than one such spot is
(pyridin-2-yl)propan-1-amine hydrogen maleate. more intense than the spot in the chromatogram obtained
with reference solution (b).
Pheniramine Maleate contains not less than 99.0 per cent and
not more than 101.0 per cent of C16H20N2,C4H4O4, calculated Heavy metals (2.3.13). Dissolve 1.0 g in 10 ml of water and add
on the dried basis. 2 ml of acetic acid and sufficient water to produce 25 ml. The
resulting solution complies with the limit test for heavy metals,
Description. A white or almost white, crystalline powder;
Method A (20 ppm).
odourless or almost odourless.
Sulphated ash (2.3.18). Not more than 0.1 per cent.
Identification
Loss on drying (2.4.19). Not more than 0.5 per cent, determined
A. Determine by infrared absorption spectrophotometry (2.4.6). on 1.0 g by drying in an oven at 60º at a pressure not exceeding
Compare the spectrum with that obtained with pheniramine 0.7 kPa.
maleate RS or with the reference spectrum of pheniramine Assay. Weigh accurately about 0.4 g, dissolve in 20 ml of
maleate. anhydrous glacial acetic acid. Titrate with 0.1 M perchloric
B. When examined in the range 230 nm to 360 nm (2.4.7), a acid, using 1- naphtholbenzein solution as indicator. Carry
0.002 per cent w/v solution in 0.1 M hydrochloric acid shows out a blank titration.
an inflection at about 262 nm; absorbance at about 265 nm, 1 ml of 0.1 M perchloric acid is equivalent to 0.01782 g of
about 0.42. C16H20N2,C4H4O4.
C. Dissolve 0.25 g in 5 ml of water, add 2 ml of strong ammonia Storage. Store protected from light and moisture.
solution and extract with three quantities, each of 5 ml, of
chloroform. Evaporate the aqueous extract to dryness, add
0.2 ml of 1 M sulphuric acid and 5 ml of water, extract with
four quantities, each of 25 ml, of ether and evaporate the Pheniramine Injection
combined ether extracts to dryness in a current of warm air. To
Pheniramine Maleate Injection
the residue add 50 mg of resorcinol and 1 ml of sulphuric
acid, heat in a water-bath for 2 minutes, shake well, heat in a Pheniramine Injection is a sterile solution of Pheniramine
water-bath for a further 30 minutes and cool in ice. Carefully Maleate in Water for Injections.

926
IP 2007 PHENIRAMINE TABLETS

Pheniramine Injection contains not less than 90.0 per cent and Pheniramine Tablets
not more than 110.0 per cent of the stated amount of
pheniramine maleate, C16H20N2, C4H4O4. Pheniramine Maleate Tablets
Identification Pheniramine Tablets contain not less than 92.5 per cent and
not more than 107.5 per cent of the stated amount of
Determine by thin-layer chromatography (2.4.17), coating the pheniramine maleate, C16H20N2, C4H4O4.
plate with silica gel GF254.
Mobile phase. A mixture of 50 volumes of cyclohexane,
Identification
40 volumes of chloroform and 10 volumes of diethylamine. Boil a quantity of the powdered tablets containing about 0.5 g
Test solution. Evaporate an appropriate volume of the injection of Pheniramine Maleate with 150 ml of acetone under a reflux
to dryness in a current of nitrogen using the minimum amount condenser for about 45 minutes. Filter and evaporate the filtrate
of heat, dissolve the residue in sufficient chloroform to to dryness on a water-bath. The residue complies with the
produce a solution containing 2.0 per cent w/v solution of following tests.
Pheniramine Maleate and centrifuge. A. Determine by infrared absorption spectrophotometry (2.4.6).
Reference solution. A 2.0 per cent w/v solution of pheniramine Compare the spectrum with that obtained with pheniramine
maleate RS in chloroform. maleate RS or with the reference spectrum of pheniramine
Apply to the plate 10 µl of each solution. After development, maleate.
dry the plate in air and examine in ultraviolet light at 254 nm. B. When examined in the range 230 nm to 360 nm (2.4.7), a
The two principal spots in the chromatogram obtained with 0.002 per cent w/v solution in 0.1 M hydrochloric acid shows
the test solution correspond to those in the chromatogram an inflection at about 262 nm; absorbance at about 265 nm,
obtained with the reference solution. Spray the plate with about 0.42.
dilute potassium iodobismuthate solution. The principal spot C. Dissolve 0.25 g in 5 ml of water, add 2 ml of strong ammonia
in the chromatogram obtained with the test solution solution and extract with three quantities, each of 5 ml, of
corresponds to that in the chromatogram obtained with the chloroform. Evaporate the aqueous extract to dryness, add
reference solution. 0.2 ml of 1 M sulphuric acid and 5 ml of water, extract with
Tests four quantities, each of 25 ml, of ether and evaporate the
combined ether extracts to dryness in a current of warm air. To
pH (2.4.24). 4.5 to 5.5. the residue add 50 mg of resorcinol and 1 ml of sulphuric
Related substances. Determine by the method described under acid, heat in a water-bath for 2 minutes, shake well, heat in a
the Identification test using as the reference solution a water-bath for a further 30 minutes and cool in ice. Carefully
solution prepared by diluting 1 volume of the test solution to add 5 ml of water; a yellow colour is produced. To 2 ml of the
500 volumes with chloroform. Any secondary spot in the solution add 3 ml of a 50 per cent w/v solution of ammonium
chromatogram obtained with the test solution is not more acetate, previously cooled in ice; a pink colour is produced
intense than the spot in the chromatogram obtained with the which persists for at least 10 minutes in the cooled solution.
reference solution.
Tests
Other tests. Complies with the tests stated under Parenteral
Preparations (Injections). Related substances. Determine by thin-layer chromatography
Assay. To an accurately measured volume containing about (2.4.17), coating the plate with silica gel G.
0.11 g of Pheniramine Maleate add sufficient water to produce Mobile phase. A mixture of 50 volumes of cyclohexane,
50.0 ml and mix well. To 20.0 ml add sufficient 1 M sodium 40 volumes of chloroform and 10 volumes of diethylamine.
hydroxide to make the solution just alkaline to litmus paper, Test solution. Shake a quantity of the powdered tablets
add 2 ml in excess and extract with two quantities, each of containing 20 mg of Pheniramine Maleate with 10 ml of
50 ml, of ether. Wash each ether extract in succession with 20, methanol, centrifuge and use the supernatant liquid.
20 and 5 ml of 0.1 M hydrochloric acid, dilute the combined
extracts to 100.0 ml with 0.1 M hydrochloric acid and mix. Reference solution (a). Dilute 1 volume of the test solution to
Dilute 5.0 ml to 100.0 ml with 0.1 M hydrochloric acid and 100 volumes with methanol.
measure the absorbance of the resulting solution at the Reference solution (b). Dilute 1 volume of reference solution
maximum at about 265 nm (2.4.7), using 0.1 M hydrochloric (a) to 20 volumes with methanol.
acid as the blank. Calculate the content of C16H20N2, C4H4O4 Apply to the plate 10 µl of each solution. After development,
taking 210 as the specific absorbance at 265 nm. dry the plate in air, spray with dilute potassium
Storage. Store protected from light. iodobismuthate solution. Any secondary spot in the

927
PHENOBARBITONE IP 2007

chromatogram obtained with the test solution is not more difference between the melting points, which are about 175º,
intense than the spot in the chromatogram obtained with is not greater than 2º.
reference solution (a) and not more than one such spot is C. Complies with the test for identification of barbiturates
more intense than the spot in the chromatogram obtained (2.3.2).
with reference solution (b).
D. Dissolve about 20 mg in 5 ml of ethanol, add a drop of
Other tests. Complies with the tests stated under Tablets. cobalt chloride solution and a drop of dilute ammonia
Assay. Weigh and powder 20 tablets. Weigh accurately a solution; a violet colour is produced.
quantity of the powder containing about 45 mg of Pheniramine E. Gives the reaction of non-nitrogen substituted barbiturates
Maleate, shake with 20 ml of 0.1 M hydrochloric acid, centrifuge (2.3.1).
and transfer the supernatant liquid to a 100-ml volumetric flask.
Repeat the extraction with three further quantities, each of 20 Tests
ml, of 0.1 M hydrochloric acid. Combine the extracts and add
Appearance of solution. A 10.0 per cent w/v solution in a
sufficient 0.1 M hydrochloric acid to produce 100.0 ml. Mix
mixture of 20 volumes of 2 M sodium hydroxide and 30 volumes
and dilute 5.0 ml to 100.0 ml with 0.1 M hydrochloric acid;
of water is clear (2.4.1), and not more intensely coloured than
measure the absorbance of the resulting solution at the
reference solution YS6 (2.4.1).
maximum at about 265 nm (2.4.7), using 0.1 M hydrochloric
acid as the blank. Calculate the content of C16H20N2,C4H4O4 Acidity. Mix 1.0 g with 50 ml of water, boil for 2 minutes, allow
taking 210 as the specific absorbance at 265 nm. to cool, filter and adjust the volume to 50 ml. To 10 ml of the
filtrate add 0.15 ml of methyl red solution; not more than 0.1 ml
Storage. Store protected from light and moisture.
of 0.1 M sodium hydroxide is required to change the colour of
the solution from orange-yellow to pure yellow.
Related substances. Complies with the test for related
Phenobarbitone substances in barbiturates (2.3.4).
Phenobarbital Sulphated ash (2.3.18). Not more than 0.1 per cent.

H Loss on drying (2.4.19). Not more than 1.0 per cent w/w,
O N O determined on 1.0 g by drying in an oven at 105º for 2 hours.
H3C Assay. Weigh accurately about 0.1 g, dissolve in 5 ml of
NH pyridine, add 0.25 ml of thymolphthalein solution and 10 ml
of silver nitrate-pyridine reagent and titrate with 0.1 M
O
ethanolic sodium hydroxide until a pure blue colour is
obtained. Repeat the operation without the substance under
C12H12N2O3 Mol. Wt. 232.2 examination. The difference between the titrations represents
Phenobarbitone is 5-ethyl-5-phenylbarbituric acid. the amount of sodium hydroxide required.
1 ml of 0.1 M ethanolic sodium hydroxide is equivalent to
Phenobarbitone contains not less than 99.0 per cent and not
0.01161 g of C12H12N2O3.
more than 101.0 per cent of C12H12N2O3, calculated on the
dried basis. Storage. Store protected from moisture.
Description. Colourless crystals or a white, crystalline powder;
odourless.
Phenobarbitone Sodium
Identification Phenobarbital Sodium; Soluble Phenobarbitone; Soluble
Test A may be omitted if tests B, C, D and E are carried out. Phenobarbital
Tests C, D and E may be omitted if tests A and B are carried
out. H
O N ONa
A. Determine by infrared absorption spectrophotometry (2.4.6). H3C
Compare the spectrum with that obtained with phenobarbitone N
RS or with the reference spectrum of phenobarbitone.
O
B. Determine the melting point (2.4.21) of the substance under
examination and of a mixture of equal quantities of the
substance under examination and phenobarbitone RS. The C12H11N2NaO3 Mol. Wt. 254.2

928
IP 2007 PHENOBARBITONE INJECTION

Phenobarbital Sodium; Soluble Phenobarbitone; Soluble Test solution. A 1 per cent w/v solution of the substance
Phenobarbital. under examination in ethanol (50 per cent).
Phenobarbitone Sodium contains not less than 99.0 per cent Reference solution. A 0.005 per cent w/v solution of the
and not more than 101.0 per cent of C12H11N2NaO3, calculated substance under examination in ethanol (50 per cent).
on the dried basis.
Loss on drying (2.4.19). Not more than 7.0 per cent, determined
Description. A white powder or crystalline granules or flaky on 0.5 g by drying in an oven at 150º for 4 hours.
crystals; hygroscopic.
Assay. Weigh accurately about 0.15 g, dissolve in 2 ml of
Identification water and add 8 ml of 0.05 M sulphuric acid. Heat to boiling
and cool. Add 30 ml of methanol and shake until dissolution
Test A may be omitted if tests B, C, D, E and F are carried out. is complete. Titrate with 0.1 M sodium hydroxide, determining
Tests C, D and E may be omitted if tests A, B and F are carried the end-point potentiometrically (2.4.25). After the first
out. inflection, stop the addition of the sodium hydroxide, add
A. Dissolve 0.2 g in 20 ml of ethanol (50 per cent), acidify 10 ml of pyridine, mix and continue the titration until the
with dilute hydrochloric acid and extract with 50 ml of ether. second inflection is reached. The difference between the
Wash the ether layer with 10 ml of water, dry over anhydrous volumes represents the amount of sodium hydroxide
sodium sulphate and filter. Evaporate the filtrate to dryness required.
and dry the residue at 105º.
1 ml of 0.1 M sodium hydroxide is equivalent to 0.02542 g of
On the residue, determine by infrared absorption C12H11N2NaO3.
spectrophotometry (2.4.6). Compare the spectrum with that
Storage. Store protected from moisture.
obtained with phenobarbitone RS or with the reference
spectrum of phenobarbitone.
B. Determine the melting point (2.4.21), of the residue obtained
in test A and of a mixture of equal quantities of the residue and
phenobarbitone RS. The difference between the melting
Phenobarbitone Injection
points, which are about 175º, is not greater than 2º. Phenobarbital Sodium Injection; Phenobarbitone Sodium
C. Complies with the test for identification of barbiturates Injection; Soluble Phenobarbitone Injection
(2.3.2), but using the following solutions. Phenobarbitone Injection is a sterile solution of
Test solution. A 0.1 per cent w/v solution of the substance Phenobarbitone Sodium in a mixture of nine volumes of
under examination in ethanol (50 per cent). Propylene Glycol and one volume of Water for Injections.
Reference solution. A 0.09 per cent w/v solution of Phenobarbitone Injection contains not less than 95.0 per cent
phenobarbitone RS in ethanol (50 per cent). and not more than 105.0 per cent of the stated amount of
phenobarbitone sodium, C12H11N2NaO3.
D. 1 g dissolves completely in 20 ml of ethanol (90 per cent)
(distinction from barbitone sodium).
Identification
E. Gives the reaction of non-nitrogen substituted barbiturates
(2.3.1). To a volume containing 1 g of Phenobarbitone Sodium add
15 ml of water if necessary, make slightly acidic with 1 M
F. Ignite about 0.1 g; the residue gives the reactions of sodium sulphuric acid and filter. The residue, after washing with water
salts (2.3.1). and drying at 105º, complies with the following tests.
Tests A. Determine by infrared absorption spectrophotometry (2.4.6).
Appearance of solution. A 10.0 per cent w/v solution in ethanol Compare the spectrum with that obtained with phenobarbitone
(50 per cent) is clear (2.4.1), and not more intensely coloured RS or with the reference spectrum of phenobarbitone.
than reference solution YS7 (2.4.1). B. Dissolve 50 mg in 2 ml of a 0.2 per cent w/v solution of
pH (2.4.24). Not more than 10.2, determined in a 10.0 per cent cobalt acetate in methanol, warm, add 50 mg of powdered
w/v solution. borax and heat to boiling; a bluish violet colour is produced.

Related substances. Complies with the test for related Tests


substances in barbiturates (2.3.4), but using the following
solutions. pH (2.4.24).10.0 to 11.0.

929
PHENOBARBITONE SODIUM TABLETSS IP 2007

Other tests. Complies with the tests stated under Parenteral Tests
Preparations (Injections).
Other tests. Complies with the tests stated under Tablets.
Assay. Weigh accurately about 2.0 g, add 30 ml of water and
3 g of sodium carbonate, stir to dissolve and titrate with Assay. Weigh and powder 20 tablets. Weigh accurately a
0.1 M silver nitrate until a distinct turbidity is observed when quantity of the powder containing about 0.3 g of
viewed against a black background, the solution being stirred Phenobarbitone Sodium, dissolve as completely as possible
vigorously throughout the titration. in 10 ml of a 2 per cent w/v solution of sodium hydroxide,
saturate with sodium chloride, acidify with hydrochloric acid
1 ml of 0.1 M silver nitrate is equivalent to 0.02542 g of and extract with successive quantities, each of 15 ml, of ether
C12H11N2NaO3. until complete extraction is effected. Wash the combined
Determine the weight per ml of the injection (2.4.29) and extracts with two quantities, each of 2 ml, of water and extract
calculate the percentage weight in volume of C12H11N2NaO3. the combined washings with 10 ml of ether. Add the ether to
the main ether layer and dry the residue to constant weight at
Storage. Store in single dose containers. 105º.
Labelling. The label states that the injection should not be 1 g of the residue is equivalent to 1.095 g of C12H11N2NaO3.
used if the solution is discoloured or if it contains a precipitate.
Storage. Store protected from moisture.

Phenobarbitone Sodium Tablets Phenobarbitone Tablets


Phenobarbital Sodium Tablets; Soluble Phenobarbitone Phenobarbital Tablets
Tablets; Soluble Phenobarbital Tablets Phenobarbitone Tablets contain not less than 92.5 per cent
and not more than 107.5 per cent of the stated amount of
Phenobarbitone Sodium Tablets contain not less than
phenobarbitone, C12H12N2O3.
92.5 per cent and not more than 107.5 per cent of the stated
amount of phenobarbitone sodium, C12H11N2NaO3.
Identification
Identification Extract a quantity of the powdered tablets containing about
0.5 g of Phenobarbitone with 50 ml of ether, filter through
A. Heat 0.1 g of the residue obtained in Assay on a water-bath
anhydrous sodium sulphate and evaporate the ether to
with 15 ml of ethanol (25 per cent) until dissolved, filter while
dryness on a water-bath. The residue complies with the
hot and allow to cool. Filter through a sintered-glass crucible,
following tests.
wash with a small quantity of ethanol (25 per cent) and dry at
105º. Heat in a sealed tube at 105º for 1 hour. A. Determine by infrared absorption spectrophotometry (2.4.6).
Compare the spectrum with that obtained with phenobarbitone
On the residue, determine by infrared absorption
RS or with the reference spectrum of phenobarbitone.
spectrophotometry (2.4.6). Compare the spectrum with that
obtained with phenobarbitone RS or with the reference B. Dissolve 50 mg in 2 ml of a 0.2 per cent w/v solution of
spectrum of phenobarbitone. cobaltous acetate in methanol, warm, add 50 mg of powdered
borax and heat to boiling; a bluish violet colour is produced.
B. The residue obtained in test A melts at about 175º (2.4.21).
C. Dissolve 50 mg of the residue obtained in Assay in 2 ml of Tests
a 0.2 per cent w/v solution of cobaltous acetate in methanol,
warm, add 50 mg of powdered borax and heat to boiling; a Disintegration (2.5.1). 30 minutes
bluish violet colour is produced. Other tests. Complies with the tests stated under Tablets.
D. Triturate a quantity of the powdered tablets containing Assay. Weigh and powder 20 tablets. Weigh accurately a
0.2 g of Phenobarbitone Sodium with 5 ml of water and filter; quantity of the powder containing about 0.3 g of
the filtrate is alkaline to litmus solution and yields a white Phenobarbitone and extract in a continuous extraction
precipitate on the addition of dilute hydrochloric acid. apparatus (2.1.8) with ether until complete extraction is
effected. Remove the ether and dry the residue, which is
E. The powdered tablets, when moistened with hydrochloric
C12H12N2O3, to constant weight at 105º.
acid and introduced on a platinum wire into a flame, impart a
yellow colour to the flame. Storage. Store protected from moisture.

930
IP 2007 PHENOLPHTHALEIN

Phenol under examination. The difference between the titrations


represents the amount of bromine required.
Carbolic acid
1 ml of 0.05 M bromine is equivalent to 0.001569 g of C6H6O.

OH Storage. Store protected from light and moisture.

Phenolphthalein
C6H6O Mol. Wt. 94.1
O
Phenol contains not less than 99.0 per cent and not more than
100.5 per cent of C6H6O. O
Description. Colourless or faintly pink or faintly yellowish
crystals or crystalline masses; odour, characteristic;
deliquescent. OH

Identification HO
A. 0.5 g dissolves completely in 2 ml of strong ammonia
solution. Dilute this solution to about 100 ml with water and C20H14O4 Mol. Wt. 318.3
to 2 ml of the resulting solution add 0.05 ml of sodium Phenolphthalein is 3,3-bis(4-hydroxyphenyl)phthalide.
hypochlorite solution (3 per cent Cl); a blue colour develops
Phenolphthalein contains not less than 98.0 per cent and not
which becomes progressively more intense.
more than 102.0 per cent of C20H14O4, calculated on the dried
B. Dissolve 1.0 g in sufficient water to produce 15 ml (solution basis.
A) and to 1 ml add 10 ml of water and 0.1 ml of ferric chloride
Description. A white or yellowish white, crystalline or
solution; a violet colour is produced which disappears on the
amorphous powder; odourless or almost odourless.
addition of 5 ml of 2-propanol.
C. To 1 ml of solution A add 10 ml of water and 1 ml of bromine Identification
solution; a pale yellow precipitate is produced.
A. Dissolves in dilute solutions of alkali hydroxides and in
Tests hot solutions of alkali carbonates forming a red solution which
is decolorised by dilute acids.
Appearance of solution. Solution A is clear (2.4.1), and not
B. Melting range (2.4.21). 258º to 263º.
more intensely coloured than reference solution BS6 (2.4.1).
Acidity. To 2 ml of solution A add 0.05 ml of methyl orange Tests
solution; the solution is yellow.
Heavy metals (2.3.13). Heat 5 g with 50 ml of dilute
Freezing point (2.4.11). Not less than 39.5º. hydrochloric acid on a water-bath for 5 min and filter.
Non-volatile matter. Not more than 0.05 per cent, when 5.0 g is Evaporate the filtrate almost to dryness and dissolve the
volatilised on a water-bath and dried to constant weight at residue in 50 ml of water. 12 ml of this solution complies with
105º. the limit test for heavy metals, Method D (20 ppm). Use 10 ml
of lead standard solution (2 ppm Pb) to prepare the standard.
Assay. Weigh accurately about 0.5 g and dissolve in sufficient
water to produce 250.0 ml. Transfer 25.0 ml to a ground-glass- Fluoran. 0.5 g dissolves completely in a mixture of 4 ml of 1 M
stoppered flask, add 50.0 ml of 0.05 M bromine and 5 ml of sodium hydroxide and 50 ml of water.
hydrochloric acid, stopper, allow to stand for 30 minutes, Sulphated ash (2.3.18). Not more than 0.1 per cent.
swirling occasionally, and allow to stand for a further
Loss on drying (2.4.19). Not more than 1.0 per cent, determined
15 minutes. Add 5 ml of a 20 per cent w/v solution of potassium
on 1.0 g by drying in an oven at 105º.
iodide, shake and titrate with 0.1 M sodium thiosulphate until
a faint yellow colour remains. Add 0.5 ml of starch solution Assay. Weigh accurately about 0.1 g, dissolve in 100.0 ml of
and 10 ml of chloroform and continue the titration with ethanol (95 per cent), dilute 5.0 ml of this solution to 50.0 ml
vigorous shaking. Repeat the operation without the substance with ethanol (95 per cent) and evaporate 5.0 ml of the resulting

931
PHENOXYMETHYLPENICILLIN POTASSIUM IP 2007

solution to dryness on a water-bath. Dissolve the residue in Phenoxyacetic acid. (Not more than 0.5 per cent).
sufficient glycine buffer pH 11.3 to produce 100.0 ml, mix and Determine by liquid chromatography (2.4.14).
immediately measure the absorbance of the resulting solution
at the maximum at about 555 nm (2.4.7). Calculate the content Diluent. pH 6.6.phosphate buffer.
of C20H14O4 taking 1055 as the specific absorbance at 555 nm. Test solution. Weigh accurately a suitable quantity of the
Storage. Store protected from moisture. substance under examination, dissolve in the diluent and dilute
to obtain a solution having a known concentration of about
20.0 mg per ml. (Use this solution on the day prepared).
Reference solution. Weigh accurately a suitable quantity of
Phenoxymethylpenicillin Potassium phenoxyacetic acid, dissolve in the diluent and dilute to obtain
Penicillin V Potassium a solution having a known concentration of about 0.1 mg per
ml.
H COOK Chromatographic system
O
O N CH3 – a stainless steel column 25 cm x 4.6 mm, packed with
O octadecylsilane chemically bonded to porous silica
N S CH3 (5 µm),
H H H – mobile phase: a mixture of 65 volumes of water,
35 volumes of acetonitrile and 1 volume of glacial acetic
acid,
C16H17KN2O5S Mol. Wt. 388.5
– flow rate. 1 ml per minute,
Phenoxymethylpenicillin Potassium is potassium (6R)-6-(2- – spectrophotometer set at 254 nm,
phenoxyacetamido)penicillinate, produced by the growth of – a 20 µl loop injector.
certain strains of Penicillium notatum or related organisms
Inject the reference solution. The tailing factor is not more
on a culture medium containing an appropriate precursor, or
than 1.5 and the relative standard deviation for replicate
obtained by any other means.
injections is not more than 2.0 per cent.
Phenoxymethylpenicillin Potassium contains not less than
Inject alternately the test solution and the reference solution.
86.0 per cent of total penicillins C16H17N2O5S, calculated on
the anhydrous basis. Calculate the content of phenoxyacetic acid.
Description. A white, crystalline powder. Limit of p-hydroxyphenoxymethylpenicillin. Not more than
5.0 per cent.
Identification Using the chromatograms obtained with the test solution in
Test A may be omitted if tests B and C are carried out. Test B the Assay, calculate the content of p-hydroxyphenoxymethyl-
may be omitted if tests A and C are carried out. penicillin from the peak response of p-hydroxyphenoxy-
methylpenicillin and the sum of the peak responses of
A. Determine by infrared absorption spectrophotometry (2.4.6). p-hydroxyphenoxymethylpenicillin and phenoxymethyl-
Compare the spectrum with that obtained with penicillin.
phenoxymethylpenicillin potassium RS.
Water (2.3.43). Not more than 1.0 per cent, determined on
B. Gives reaction B of penicillins and cephalosporins (2.3.1). 1.0 g.
C. Gives reaction A of potassium salts (2.3.1). Assay. Determine by liquid chromatography (2.4.14).

Tests Test solution. Dissolve about 125 mg of the substance under


examination in the mobile phase and dilute to 50.0 ml with the
pH (2.4.24). 5.5 to 7.5, determined in a 0.5 per cent w/v solution. mobile phase.
Specific optical rotation (2.4.22). +215.0º to +230.0º, determined Reference solution (a). Dissolve an accurately weighed
in a 1.0 per cent w/v solution in carbon dioxide-free water. quantity of phenoxymethylpenicillin potassium RS in the
mobile phase and dilute to obtain a solution having a known
Light absorption (2.4.7). Absorbance of a 0.1 per cent w/v
concentration of about 2.5 mg per ml.
solution in 0.1 M sodium hydroxide at the maximum at about
306 nm, not more than 0.33. Absorbance of a 0.02 per cent Reference solution (b). A solution in the mobile phase
w/v solution in 0.1M sodium hydroxide at the maximum at containing 0.25 per cent w/v each of benzylpenicillin potassium
about 274 nm, not less than 0.50. and phenoxymethylpenicillin potassium.

932
IP 2007 PHENOXYMETHYLPENICILLIN POTASSIUM TABLETS

Chromatographic system add 1.0 ml of penicillinase solution; the solution changes


– a stainless steel column 30 cm x 4 mm, packed with rapidly to red.
octadecylsilane chemically bonded to porous silica C. Ignite 0.5 g of the powdered tablets, add 5 ml of 2 M
(3 to 10 µm), hydrochloric acid, boil, cool and filter. The filtrate gives
– mobile phase: a mixture of 650 volumes of water, reaction B of potassium salts (2.3.1).
350 volumes of acetonitrile and 5.75 volumes of glacial
acetic acid, Tests
– flow rate. I ml per minute,
Dissolution (2.5.2).
– spectrophotometer set at 254 nm,
– a 10 µl loop injector. Apparatus. No 1
Medium. 900 ml of water
Inject reference solution (b).The relative retention times are
Speed and time. 50 rpm and 45 minutes.
about 0.8 for benzylpenicillin and 1.0 for phenoxymethyl-
penicillin. The column efficiency determined from the Withdraw a suitable volume of the medium and filter. Measure
phenoxymethylpenicillin peak is not less than the absorbance of the filtrate, suitably diluted if necessary, at
1800 theoretical plates and the resolution between benzyl- the maximum at about 268 nm (2.4.7). At the same time measure
penicillin and phenoxymethylpenicillin is not less than 3.0. the absorbance of a solution of known concentration of
phenoxymethylpenicillin potassium RS at the maximum at
Inject reference solution (a). The relative standard deviation
about 268 nm. Calculate the content of C16H18N2O5S, in the
for replicate injections is not more than 1.0 per cent.
medium.
Inject the test solution and reference solution (a). Record the
D. Not less than 75 per cent of the stated amount of
chromatograms and measure the responses for the
C16H18N2O5S.
phenoxymethylpenicillin peak and any p-hydroxyphenoxy-
methylpenicillin peak with a retention time of about 0.4 relative Other tests. Complies with the tests stated under Tablets.
to that of the main phenoxymethylpenicillin peak. Assay. Determine by liquid chromatography (2.4.14).
Calculate the content of C16H17N2O5S, from the sum of the Test solution. Weigh and finely powder 20 tablets. Dissolve
peak responses of the p-hydroxyphenoxymethylpenicillin and an accurately weighed quantity of the powder containing about
phenoxymethylpenicillin peaks in the chromatograms obtained 0.25 g of phenoxymethylpenicillin in the mobile phase by
with the test solution and reference solution (a). shaking for 5 minutes and dilute to 100.0 ml with the mobile
Storage. Store protected from moisture. phase. Filter through a 0.5 µm or finer filter and use the filtrate.
Reference solution (a). Dissolve an accurately weighed
quantity of phenoxymethylpenicillin potassium RS in the
mobile phase and dilute to obtain a solution having a known
Phenoxymethylpenicillin Potassium concentration of about 2.5 mg per ml.
Tablets Reference solution (b). A solution in the mobile phase
Phenoxymethylpenicillin Tablets; Penicillin V Potassium containing 0.25 per cent w/v each of benzylpenicillin potassium
Tablets; Penicillin V Tablets and phenoxymethylpenicillin potassium.
Chromatographic system
Phenoxymethylpenicillin Potassium Tablets contain not less
– a stainless steel column 30 cm x 4 mm, packed with
than 92.5 per cent and not more than 107.5 per cent of the
octadecylsilane chemically bonded to porous silica,
stated amount of phenoxymethylpenicillin, C16H18N2O5S.
– mobile phase: a mixture of 650 volumes of water,
Identification 350 volumes of acetonitrile and 5.75 volumes of glacial
acetic acid,
A. Shake a quantity of the powdered tablets containing 80 mg – flow rate. 1 ml per minute,
of phenoxymethylpenicillin with water, dilute to 250 ml with – spectrophotometer set at 254 nm,
water and filter. When examined between 230 and 360 nm – a 10 µl loop injector.
(2.4.7), the filtrate shows absorption maxima at about 268 nm
Inject reference solution (b).The relative retention times are
and 274 nm and a minimum at about 272 nm.
about 0.8 for benzylpenicillin and 1.0 for phenoxymethyl-
B. Shake a quantity of the powdered tablets containing 10 mg penicillin. The column efficiency determined from the
of phenoxymethylpenicillin with 10 ml of water, filter and add phenoxymethylpenicillin peak is not less than 1800 theoretical
0.5 ml of neutral red solution. Add sufficient 0.01 M sodium plates and the resolution between the benzylpenicillin and
hydroxide to produce a permanent orange colour and then phenoxymethylpenicillin peaks is not less than 3.0.

933
PHENOTOLAMINE MESYLATE IP 2007

Inject reference solution (b). The relative standard deviation D. Mix 50 mg with 0.2 g of powdered sodium hydroxide, heat
for replicate injections is not more than 1.0 per cent. to fusion and continue the heating for a few seconds longer.
Inject the test solution and reference solution (a). Record the Cool, add 0.5 ml of water and a slight excess of 2 M
chromatograms and measure the responses for the hydrochloric acid and warm; sulphur dioxide is evolved, which
phenoxymethylpenicillin peak and any p-hydroxyphenoxy- turns moistened starch iodate paper blue.
methylpenicillin peak with a retention time of about 0.4 relative
to that of the main phenoxymethylpenicillin peak.
Tests

Calculate the content of C16H17N2O5S, from the sum of the Acidity or alkalinity. Dissolve 0.1 g in 10 ml of carbon dioxide-
peak responses of the p-hydroxyphenoxymethylpenicillin and free water and add 0.1 ml of methyl red solution. The solution
phenoxymethylpenicillin peaks in the chromatograms obtained is not red and not more than 0.05 ml of 0.1 M sodium hydroxide
with the test solution and reference solution (a) is required to change the colour of the solution.
Storage. Store protected from moisture. Related substances. Determine by thin-layer chromatography
(2.4.17), coating the plate with silica gel G.
Labelling. The label states the strength in terms of the
equivalent amount of phenoxymethylpenicillin. Mobile phase. A mixture of 85 volumes of 2-butanone,
15 volumes of acetone and 5 volumes of strong ammonia
solution.

Phentolamine Mesylate Test solution. Dissolve 0.2 g of the substance under


examination in 10 ml of ethanol (95 per cent).
OH Reference solution. A 0.01 per cent w/v solution of the
CH3 substance under examination in ethanol (95 per cent).
Apply to the plate 10 µl of each solution. After development,
N , CH3SO3H dry the plate in air, spray with dilute potassium
N iodobismuthate solution. Any secondary spot in the
chromatogram obtained with the test solution is not more
HN intense than the spot in the chromatogram obtained with the
reference solution.
C17H19N3O,CH4O3S Mol. Wt. 377.5
Sulphated ash (2.3.18). Not more than 0.1 per cent.
Phentolamine Mesylate is 3-[[4,5-dihydro-1H-imidazol-2-
yl)methyl](4-methylphenyl)aminophenol Loss on drying (2.4.19). Not more than 0.5 per cent, determined
methanesulphonate. on 1.0 g by drying in an oven at 105º.
Phentolamine Mesylate contains not less than 99.0 per cent Assay. Weigh accurately about 0.3 g and dissolve in 100 ml of
and not more than 100.5 per cent of C17H19N3O, CH4O3S, anhydrous 2-propanol with the aid of ultrasound if necessary.
calculated on the dried basis. Titrate with 0.1 M tetrabutylammonium hydroxide in
2-propanol. Determine the end-point potentiometrically
Identification (2.4.25), using a glass electrode and a calomel electrode
Test A may be omitted if tests B, C and D are carried out. Tests containing a saturated solution of tetramethylammonium
B, C and D may be omitted if test A is carried out. chloride in 2-propanol. Carry out a blank titration.

A. Determine by infrared absorption spectrophotometry (2.4.6). 1 ml of 0.1 M tetrabutylammonium hydroxide is equivalent to


Compare the spectrum with that obtained with phentolamine 0.03775 g of C17H19N3O, CH4O3S.
mesylate RS or with the reference spectrum of phentolamine Storage. Store protected from light and moisture.
mesylate.
B. When examined in the range 230 nm to 360 nm (2.4.7), a
0.002 per cent w/v solution shows an absorption maximum
only at about 278 nm; absorbance at about 278 nm, about 0.5. Phentolamine Injection
C. Dissolve 0.5 g in 5 ml of ethanol (95 per cent) and 5 ml of Phentolamine Mesylate Injection
0.1 M hydrochloric acid and add 2 ml of a 0.5 per cent w/v
solution of ammonium metavanadate; a light green precipitate Phentolamine Injection is a sterile solution of Phentolamine
is produced. Mesylate in Water for Injections containing Dextrose.

934
IP 2007 PHENYLBUTAZONE

Phentolamine Injection contains not less than 95.0 per cent B. When examined in the range 230 nm to 360 nm (2.4.7), a
and not more than 105.0 per cent of the stated amount of 0.001 per cent w/v solution in 0.01 M sodium hydroxide shows
phentolamine mesylate, C17H19N3O, CH4O3S. an absorption maximum at about 264 nm; absorbance at about
264 nm, about 0.66.
Identification
C. To 0.1 g add 1 ml of glacial acetic acid and 2 ml of
The residue obtained in the Assay melts at about 138º (2.4.21). hydrochloric acid and heat under a reflux condenser for 30
minutes. Cool, add 10 ml of water and filter. Add to the filtrate
Tests 3 ml of 0.1 M sodium nitrite; a yellow colour