CERTIFICATE

This is to certify that Ms. Priti Suman, student of JIWAJI UNIVERSITY, Gwalior,
has worked on the project titled Microbial limit test of non-sterile
products and pharmaceuticals water under my supervision and
guidance for three months (18th February-18th May 2015) at the Ind-Swift
Limited, (Baddi) Himachal Pradesh.
This dissertation is being submitted by the student to
the JIWAJI UNIVERSITY, Gwalior in fulfilment of the requirement for
the degree of Master of Science (Microbiology). No part of this
project training report has been submitted for any degree or diploma
to any other university/institute of India or Abroad.

Ms. Priti Suman (Candidate)

Dr. Vinod (Q.C Manger)

D.S Mehra (Plant Head)

ACKNOWLEGMENT

I, Priti Suman Student of M.Sc. (Microbiology) IV semester in Jiwaji

University, Gwalior (M.P) take this opportunity of expressing my
deep sense of gratitude and indebtness to D.S Mehra, plant head, Ind
Swift Limited for the suggestion and guidance in an unabated manner.
Also I am very grateful to Dr. Vinod, QC Manager, Ind Swift Limited
for her meticulous guidance and support throught out the training
tenure.
I Would also like to thank Ms. Veena yangpha, Mr. Sanjeev
for their valuable and encouraging suggestions. I will like to
acknowledge the care & support during the training tenure for their
generous help at each and every step.
I am enormously thankful to Dr. Avinsh Tiwari (Coordinator,
S.O.S in Microbiology, Jiwaji University),
(Faculty of S.O.S in
Microbiology, Jiwaji University) for their proper guidance & valuable
suggestions for my project training.
Iam also thankful to my lab mates omvir & vishal for their
help & cooperation.
I would also like to thank my parents who made me capable
for this work and made me what I am. At last but not the less I am
thankful to almighty god for his blessings on me during the time of
training and preparation of this report.
Priti Suman
M.Sc. Microbiology
Jiwaji University, Gwalior (M.P.)

CONTENTS
1.
2.
3.
4.
5.
6.
7.

Company Profile
Introduction
Review of Literature
Materials And Methods
Results And Observations
Instruments Used
References

CHAPTER 1

COMPANY PROFILE

Company Profile( Ind swift limited)

Swift is the fastest flying bird on earth. It is the philosophy behind the genesis of the name Ind-Swift
Ltd Incepted in 1986 when three visionaries the Mehtas, the Munjals and the Jains, all 1st generation
entrepreneurs visualized the business. Even with limited resources, the vision was to develop a
Pharma enterprise with its body spread internationally and soul rooted in ethics. Today with strong
business acumen, apart from pharmaceutical business Ind-swift has also progressively forayed in
diversification into multifarious fields viz Infrastructure, Printing & Packaging and Stationary,
Education, and Media Publication with its every unit as an independent profit earning centre. It is
indeed a proud moment for us today, starting as a small domestic company in 1986 we have
transformed into a truly global organisation with its operations and product range in more than 50

countries. Indswift today is 2000 Crore group, moving forward and still growing to new dimensions
and spheres every day.
Established in 1986 by the Jains, Mehtas and Munjals, Ind-Swift is a leading pharmaceutical
manufacturing and marketing company in India, based in Chandigarh. Its strength lies in innovative
pharmaceutical products. Ind-Swift has been ranked 35th in the Indian pharma industry and is the
second largest among the drug manufacturers in India (north India). It has spread its network across
45 countries. It is an ISO 9001-2008, WHO GMP certified company. It is also listed on the Bombay
Stock Exchange and National Stock Exchange. It has 5 plants in India which include multi-purpose,
multi-location facilities spread across northern India. The facilities are built according to the current
guidelines of MHRA, EU, WHO and accreditations with ISO 14000 series standards.

The company has world class expertise in finished goods dosage, Active Pharmaceutical Ingredients
(APIs) and herbal products. A talented team of research scientists, formulation experts, clinical
development and regulatory affairs professionals support the company's marketing efforts not only
across the country but also around the globe. Ind-Swift pharmaceutical products are safe, effective
and have consistent quality.
IND-SWIFT is Chandigarh based pharmaceutical company, established in 1986 with a mission of
winning global customers through innovative pharmaceutical products. Three visionaries Jains,
Mehtas and Munjals, dedicated themselves to work for humanitys quest for longer, happier and
healthier lives.
Ind-swift is a research driven forward looking pharmaceutical company with world class expertise in
finished goods dosage and active pharmaceutical ingredients (APIs)and herbal products Ind-swift is
ISO 9001-2008, WHO GMP certified and is listed on Bombay Stock Exchange and National Stock

Exchange. Ind-swift has been ranked 35th among top Indian Pharmaceutical companies. At Ind-swift
we ensure value for money and customer satisfaction globally , in doing so we have delivered long
term profitable returns to our investors, value to our partners and rewarding careers to our employees.
Ind-swift is proud to be the second largest drug manufacturers of North India. Our multipurpose;
multiplication manufacturing set-ups are spread across the lush-green plains of northern India. The
facilities are built according to current guidelines of MHRA, EU, WHO, and accreditations with ISO
14000

The company has dedicated research and development department well equipped with the latest
equipments and supported by a large pool, of scientists who continuously work towards new pharma
products.

The company possess portfolio of 750 products with presence in high growth therapeutic segments of
Cardiology, Diabetology, Anti depressant, anti-allergic, Anti- infective, Neurology & Oncology with a
nationwide distribution network. Apart from being among top Indian pharma companies, Ind-swift
has also progressively embarked in diversification into multifarious fields viz infrastructure, printing
packaging and stationery, education, and media publication with its every unit as an independent
profit earning centre.
Life is the most precious thing on earth, to serve the same, we at Ind-swift are committed to give our
unflinching efforts, support and dedication for the service of humanity by developing innovative,
therapies and process to produce safe effective and consistent quality pharmaceutical products
delivering across the globe hence making healthier and happier world "Because Life is precious".

Values :
Winning global customers through innovative pharmaceutical products.
Consistent profitable performance.

Innovation to nurture good partnership with customers.

Responsibility towards environment , safety & health.
Commitment towards highest standards of ethics & integrity.
Valuing the ability to excel, integrity, knowledge, skills, diversity and
teamwork of employees.
FACILITIES :
A unique and unmatched excellence in pharmaceuticals manufacturing with
highest reliance of product
quality attributes is our corporate strength . Ind-Swift's multipurpose,
multilocation manufacturing set-ups are
Across spread across the lush-green plains of northern Indian states viz
Himachal Pradesh, Haryana and
Jammu and Kashmir. The locations are environmental pollution free and
together offer approx.12 lacs sq. ft
of newly constructed plants with state of the art facilities for pharma
manufacturing.
The facilities are built according to current guidelines of USFDA, MHRA, EU,
and WHO, and accreditations
with ISO 14000 series standard.

A strong and ever vigilant Quality Assurance System forms the backbone of
our continuing compliance to
stringent regulatory guidelines, backed by latest equipments and
instrumentation system. We achieve
constant compliance to product quality.attributes with an enviable cost of
efficiency. It is always our strong
endeavor to continuously invest in newer products, process technologies and
look out for new horizons in
pharmaceutical business to establish our well earned leadership.

CHAPTER 2
INTRODUCTION

INTRODUCTION
Pharmaceutical Microbiology is an applied branch of Microbiology. It involves the study
of microorganisms associated with the manufacture of pharmaceuticals e.g. minimizing the
number of microorganisms in a process environment, excluding microorganisms and
microbial by-products like exotoxin and endotoxin from water and other starting materials,
and ensuring the finished pharmaceutical product is sterile. Other aspects of pharmaceutical
microbiology include the research and development of anti-infective agents, the use of
microorganisms to detect mutagenic and carcinogenic activity in prospective drugs, and the
use of microorganisms in the manufacture of pharmaceutical products like insulin and human
growth hormone.

For experienced testing of raw materials and other nonsterile products in pharmaceutical
manufacturing, turn to Microtest. We offer excellent solutions for USP Microbial Limits
Testing (MLT).
The Quantitative phase of testing determines the bioburden of given pharmaceutical
manufacturing samples, determining the number of total aerobic organisms as well as the
total number of yeasts, and molds.
The qualitative phase of testing examines samples per U.S. FDA criteria for objectionable
organisms. For minimum coverage, these criteria require testing for the following:

The bacteria Escherichia coli (E. coli)

The bacteria Staphylococcus aureus (S. aureus)

The bacteria salmonella

The bacteria Pseudomonas aeruginosa (Ps. aeruginosa)

The bacteria shigella

The first step in Microbial limits testing is the preparatory testing which validates the product
with the method used. This ensures the product has no inhibitory effect on the recovery of
organisms from the product. Once the product has passed the Preparatory Test, the product is
validated for routine screening. The routine Microbial limits screening test consists of the
total aerobic and total yeast and mold counts as well as the test for objectionable
microorganisms. If the total aerobic and total yeast and mold counts are acceptable and no
objectionable organisms are found, Microtest then gives the go-ahead for swift release of
the product or raw material.

Full microbial limits testing then inoculates specific organisms into general nutrient media.
After allowing time for any growth, our microbiologists transfer the results to specific media
that show differential reactions (usually via color changes) to the presence of a given
organism.
The Preparatory and the routine microbial limits screening test can be particularly
problematic for inexperienced laboratory personnel. With an average of more than 10 years
experience, Microtest specialists bring the industrys highest levels of expertise to the task.
Drug safety is a major focus of pharmaceutical microbiology. Pathogenic bacteria,
yeasts, moulds and toxins produced by microorganisms are all possible

contaminants of medicines- although stringent, regulated processes are in place

to ensure the risk is minimal.

Nonsterile pharmaceuticals are not produced by aseptic processes and, therefore, are not
expected to be totally free from microbial contaminations. The degree of contamination in
nonsterile products is regulated, and is based on the acceptance criteria for microbiological
quality established in Pharmacopeial monographs. A review of the U.S. Food and Drug
Administration's (FDA) enforcement reports during 20042011 revealed that approximately
75% of nonsterile product recalls were in fact due to contaminated over-the-counter
(OTC) or personal care products.
Microbial limits testing is performed to determine whether a product complies with
compendial specifications for microbial quality. This testing consists of two parts. The
quantitative phase, Microbial Enumeration, gives the total number of aerobic organisms as
well as a total yeast and mold count on a product. The qualitative phase is known as the Test
for Specified Microorganisms. This test is designed to determine the presence or absence of
specific objectionable organisms in a product.
Prior to performing MLT testing on a product, the method must be validated to ensure that the
product has no microbial inhibitory properties which could produce false negatives. This
validation testing is known as the MLT Method Suitability Test.
Microbial limits testing is performed to determine whether a product complies with
compendial specifications for microbial quality. This testing consists of two parts. The
quantitative phase, Microbial Enumeration, gives the total number of aerobic organisms as
well as a total yeast and mold count on a product. The qualitative phase is known as the Test
for Specified Microorganisms. This test is designed to determine the presence or absence of
specific objectionable organisms in a product.
Prior to performing MLT testing on a product, the method must be validated to ensure that the
product has no microbial inhibitory properties which could produce false negatives. This
validation testing is known as the MLT Method Suitability Test.

MLT Services

MLT Method Suitability Test

Microbial Enumeration (Total Aerobic Microbial Count and Total Combined Yeasts
and Mold Count)

Screening Test for Specified Microorganisms:

o P. aeruginosa
o E. coli
o S. aureus
o Salmonella
o Bile tolerant gram negative bacteria
o Shigella
o Clostridium

Water is widely used as a raw material, ingredient, and a solvent in the processing,
formulation, and manufacture of pharmaceutical products, active pharmaceutical ingredients
and intermediates, compendial articles, and analytical reagents. As such, all water purification
systems must be monitored regularly to verify the quality of the water produced. This
includes chemical purity as well as microbiological quality.
Microbiological quality is monitored through various testing processes including total
heterotrophic plate count, coliforms/E. coli monitoring, or by checking for the presence of
other organisms suspected to be present in a water sample. Chemical purity of water samples
can be assessed quantitatively through the use of Total Organic Carbon (TOC) testing.
Some types of water must also be tested for endotoxins, for example LAL reagent water, USP
Water for Injection, and water for hemodialysis. Some steam systems also require
certification of low endotoxin levels.
Water Testing Services

Total Heterotrophic Plate Count

Coliform Test / E. coli

Total Organic Carbon

LAL (Kinetic Chromogenic method)

Conductivity

pH

Limits used in criteria should be based on microbiological data appropriate to the food and
should be applicable to a variety of similar products. They should therefore be based on data
gathered at various production establishments operating under Good Hygienic Practices and
applying the HACCP system.
In the establishment of microbiological limits, any changes in the microflora likely to occur
during storage and distribution (e.g. decrease or increase in numbers) should be taken into
account.
Microbiological limits should take into consideration the risk associated with the
microorganisms, and the conditions under which the food is expected to be handled and
consumed. Microbiological limits should also take account of the likelihood of uneven
distribution of microorganisms in the food and the inherent variability of the analytical
procedure.
If a criterion requires the absence of a particular microorganism, the size and number of the
analytical unit (as well as the number of analytical sample units) should be indicated.
Quality control test for Nonsterile pharmaceutical product includes the microbiological
testing of raw materials, excipients, active ingredients, bulk, and finished products. Nonsterile
samples contain high numbers of microbes and objectionable MOs that change the chemical
composition by spoilage, affecting the stability and integrity of the product and package.
Microbial bioburden is allowed based upon the product specifications. Quantity and types of
MO will determine the safety of that particular pharmaceutical product and efficacy of the
manufacturing process.
Microbial limit test (MLT) is based upon: Chemical composition of product Production
process Route of application Intended use of product Delivery system of product USP, EP,
and JP had divided MLT into types of test Quantitative test determines number of bacteria,
yeast, and mold present in a given pharmaceutical sample and Qualitative test - determines
the presence of specific pathogen indicators, e.g., Salmonella spp., Staphylococcus aureus,
Escherichia coli, P. aeruginosa, and Enterobacteriaceae.

Salmonella and E. coli (toxin producers) are gram-negative rods , capable of lactose
fermentation , commonly found in fecal sources associated to intestinal disorders P.
aeruginosa is a gram-negative, non fermentative rod,typically associated to opportunistic
infections S. aureus is a gram positive cocci commonly associated to skin, gastrointestinal,
and toxic shock syndrome conditions.

USP recommendations and Limits: Plant, animal , and mineral -based formulations absence
of Salmonella Orally administered products - absence of E. coli Topical pharmaceutical
formulation - absence of S. aureus, P. aeruginosa Vaginal, rectal, and urethral formulations absence of yeast and mold.
EP recommendations and Limits: Tropical, transdermal, respiratory NMT 100CFUs/g or ml
of bacteria Absence of S.aureus & Pseudomonas Oral & rectal routes -NMT 1000 CFUs / g
or ml NMT 100 CFUs / g or ml of Yeast, mold Plant, animal, and mineral-based formulations
NMT 10,000 CFUs/g ml Absence of salmonella species, S.aureus, E.Coli Herbal
formulations - NMT 10 5 to 10 7 CFUs /g or ml for bacteria NMT 10 4 to 10 5 CFUs / g or
ml for yeast, mold If reconstituted with boiling water - NMT 10 2 CFUs / g or ml If boiling
water not added - Absence of E.Coli & salmonella.
MICROBIAL LIMIT TEST PREPARATORY TESTING is to be done to confirm that the
sample themselves do not inhibit multiplication of micro organisms, under the test conditions,
that may be present. diluted specimens of the material to be inoculated with separate viable
cultures of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and
Salmonella . Innoculum: 1 ml of not less than 10 -3 dilution of a 24-hour broth culture of the
microorganism in pH 7.2 Phosphate Buffer , Fluid SoybeanCasein Digest Medium , or Fluid
Lactose Medium of the test material.
Observation & Result of the preparatory test: Absence of Microbial growth in the relevant
medium invalidates the MLT and necessitates modification of the procedure by (1) an
increase in the volume of diluent , the quantity of test material remaining the same, or by (2)
addition of suitable inactivating agent in the diluents, or (3) Combination of modifications (1)
and (2) so as to permit growth of the inoculum. 0.5 % soy lecithin , and 4% polysorbate 20
are used to neutralize inhibitory substances present in the sample.
CULTURE MEDIA Culture media may be prepared as per the formula or dehydrated culture
media may be used. Dehydrated media , when reconstituted as directed by the manufacturer
or distributor, should have similar ingredients and/or yield media comparable to those
obtained from the formulas. Dissolve the soluble solids in the water, using heat, if necessary,
to effect complete solution Use Agar containing NMT 15% Moisture content. Use Purified
Water . Determine the pH at 25 2. add solutions of HCl or NaOH to adjust the pH.
Sterilization of media is by using autoclave at 121 0 C at 15psi for 15 minutes.

Culture media used in Microbial Limit test: 1.SoybeanCasein Digest Agar Medium 2.Fluid
SoybeanCasein Digest Medium 3.Pseudomonas Agar Medium for Detection of Pyocyanin
4.XyloseLysineDesoxycholate Agar Medium 5.Bismuth Sulfite Agar Medium 6.Triple
SugarIronAgar Medium 7.MacConkey Agar Medium 8.Levine EosinMethylene Blue
Agar Medium 9.Sabouraud Dextrose Agar Medium 10.Potato Dextrose Agar Medium.
10 Culture media used in Microbial Limit test: 11.Fluid Casein DigestSoy Lecithin
Polysorbate 20 Medium 12.Pseudomonas Agar Medium for Detection of Fluorescin

13. Fluid Lactose Medium 14. Fluid SeleniteCystine Medium 15. Fluid Tetrathionate
Medium 16. Brilliant Green Agar Medium 17. MannitolSalt Agar Medium 18. BairdParker
Agar Medium 19. VogelJohnson Agar Medium 20. Cetrimide Agar Medium.

Cleanrooms and controlled environments

Pharmaceutical microbiologists are required to assess cleanrooms and controlled
environments for contamination (viable and particulate) and to introduce contamination
control strategies. This includes an understanding of risk assessment.[3]
Risk management has been successfully employed in various industrial sectors like US Space
industry (NASA), nuclear power industry and automobile industry which benefited these
industries in several areas. But in application, the pharmaceutical sector is still in its infancy
and the utilization of risk assessment techniques to pharmaceutical production is just
beginning and the potential gains are yet to be realized.
Cleanrooms and zones are typically classified according to their use (the main activity within
each room or zone) and confirmed by the cleanliness of the air by the measurement of
particles. Cleanrooms are microbiologically assessed through environmental monitoring
methods.
Viable monitoring is designed to detect levels of bacteria and fungi present in defined
locations /areas during a particular stage in the activity of processing and filling a product.
Viable monitoring is designed to detect mesophilic micro-organisms in the aerobic state.
However, some manufacturers may have requirements to examine for other types of
microorganisms (such as anaerobes if nitrogen lines are used as part of the manufacturing
process)
Surface methods include testing various Surfaces for numbers of microorganisms, such as:
Product Contact Surfaces Floors Walls CeilingsUsing techniques like:
Contact Plates Touch Plates Swabs Surface Rinse Method

For air monitoring, this is undertaken using agar settle plates (placed in the locations of
greatest risk) or active (volumetric) air-samplers (to provide a quantitative assessment of the
number of microorganisms in the air per volume of air sampled). Active air-samplers
generally fall into the following different models:
Slit to Agar Membrane Filtration Centrifugal Samplers

ssMonitoring methods will all use either a general purpose culture medium like tryptone soya
agar (TSA), which will be used at a dual incubation regime of 30 C 35 C and 20 C
25 C or two different culture media are used at two different temperatures, of which one of
the media is selective for fungi (e.g. Sabouraud Dextrose agar, SDA). The choice of culture
media, incubation times and temperatures requires validating.

CHAPTER -3
REVIEW OF LITERATURE

Review of Literature
DRUG DISCOVERY

Therapeutic drugs have played a major role in increasing the average life expectancy in the
whole world in the last century. However, while many of the drugs in use in the last fifty
years or more have been of synthetic or semi-synthetic origin, the pharmacopoeias prior to
that period were of natural origin.

The medicinal value of plants has been recognized by almost every society on this planet. In
the nineteenth and earlier centuries, natural product extracts, particularly those derived from
botanical species, provided the main source of folk medicines. However, in the latter part of
the nineteenth century, biologically-active organic molecules began to be isolated in
relatively pure form for medicinal use. For example, salicyclic acid, the precursor of aspirin,
was isolated in 1874 from willow bark. Various more potent painkillers, such as morphine
and codeine, were isolated from the opium poppy.

The anti-malarial agent, quinine, was separated from cinchona (china bark). The leaves of the
purple foxglove plant provided an excellent source of digitalis that was purified for use
against heart disease. There are numerous other examples. Although synthesis of the first
synthetic pharmaceutical drug, aspirin, occurred in the latter half of the nineteenth century, it
was not until the early 1900s that the recognition of aspirin as a universal pain reliever was
realized and this discovery spawned the era of therapeutic agents.
However, it was not until the recognition that many infectious diseases were caused by
microorganisms that the real impetus in the development of therapeutic agents, both natural
and non-natural, began to occur. Concurrent with the discoveries in medical microbiology
were major advances in synthetic organic chemistry and biochemistry that provided further
momentum in the area of therapeutic agents. Synthetic sulfa drugs, the natural antibiotic,
penicillin, from Penicillium notatum , the semi-synthetic antibiotic, tetracycline, produced
from natural chlortetracycline elaborated by Streptomyces aureofaciens and the antitubercular aminoglycoside, streptomycin, from Streptomyces griseus

were all landmark

discoveries of the 1930s and 1940s. The importance of vitamins and diseases caused by their
deficiencies were also being uncovered during this period. During the next several decades
advances in X-ray crystallography, NMR spectroscopy and mass spectrometry and
developments in electrophoresis, ultracentrifugation, HPLC and other technologies
contributed to the discovery of additional chemical entities with therapeutic activities and to
the development of some vaccines. Selected examples include oral contraceptives,
tranquilizers (e.g., valium) and poliomyelitis vaccines.

The seeds for the concept of rational drug design were laid in the 1940s and 1950s by George
Hitchings and Gertrude Elion in their work on DNA-based antimetabolites, which led to the
discovery of modified purines with anticancer activity.

However, the era of DNA and medicine was largely stimulated by the elucidation of the
double-helical structure of DNA by Watson and Crick in 1953. The ramifications of this
discovery in DNA replication, transcription and translation led to a much better
understanding of viral replication. This laid the foundation for antiviral drug discovery in
subsequent decades as molecular targets in the viral replication cycle began to be identified.
The 1950s also saw the discovery of vancomycin, a glycopeptide which was developed much
later for use against methicillin-resistant staphylococci infections. The era of recombinant
DNA technology and molecular cloning began around the mid-1970s. Spectacular
accomplishments in the area of synthesis of complex, biologically-active natural products
also occurred during this period including the monumental synthesis of vitamin B12 by
Nobel Prize winner, R. B. Woodward, and his research team, which included Vasu Nair. The
natural product, taxol, isolated from Taxus brevifolia in 1971, represents an excellent
example of the combination of natural product isolation and organic synthesis in the
development of an anticancer agent. Taxol finally progressed into clinical use in 1993.
Developments in molecular biology and virology had a major impact in the scientific
understanding in the 1980s and early 1990s of the replication of the retrovirus, HIV.
Identification of the possible biochemical points of attack on the virus replication cycle gave
drug discovery efforts around the world major impetus for the design and synthesis of
nucleoside, non-nucleoside and peptide analogs as targeted anti-HIV agents.

The emergence of drug-resistant virus of both natural and non-natural origin has complicated
the therapeutic picture considerably and has necessitated the use of drug combination therapy.
The polymerase chain reaction (PCR) of the 1980s resulted in major advances in
biotechnology that have had significant impact in drug discovery.

In the 1990s, combinatorial chemistry, molecular modeling and bioinformatics contributed to

the discovery of newer generation drugs based on genomics and proteomics. The beginning
years of the new millennium will see further advances in drug discovery which will likely be
based on state-of-the art chemistry and of chemical genetics, new advances in biology,
enzyme-based molecular syntheses, proteomics and genomics, recombinant biomolecules,
high-throughput screening, and gene and cell therapy.

Water Testing
Water is widely used as a raw material, ingredient, and solvent in the processing, formulation,
and manufacture of pharmaceutical products, active pharmaceutical ingredients (APIs) and
intermediates, compendia articles, and analytical reagents. This general information chapter
provides additional information about water, its quality attributes that are not included within
a water monograph, processing techniques that can be used to improve water quality
standards that should be considered when selecting a water source.

This chapter is not an all inclusive writing on pharmaceutical water and its production meet
applicable governmental regulations, guidance, and the compendia specifications for the
types of water used in compendia articles.
Control of the chemical purity of these waters is important and is the main purpose of the
main purpose of the monographs in this compendium. Unlike other official articles, the bulk
water monographs (Purified Water & Raw Water) also limit how the article can be produced
because of the belief that the nature and robustness of the purification process is directly
related to the resulting purity.

The chemical attributes listed in these monographs should be considered as a set of minimum
specifications may be needed for some applications of these waters is found in the
monographs and is further explained in this chapter.

Purified water
Purified water is water that has been mechanically filtered or processed to remove
impurities and make it suitable for use. Distilled water has been the most common form of
purified water, but, in recent years, water is more frequently purified by other processes
including deionization (DI), reverse osmosis, carbon
filtering, microfiltration, ultrafiltration, ultraviolet oxidation, or electro-deionization.
Combinations of a number of these processes have come into use to produce water of such
high purity that its trace contaminants are measured in parts per billion (ppb) or parts per
trillion (ppt). Purified water has many uses, largely in science and engineering laboratories

and industries, and is produced in a range of purities. It can be produced on site for
immediate use or purchased in containers. Purified water in colloquial English can also refer
to water which has been treated ("rendered potable") to neutralize, but not necessarily remove
contaminants considered harmful to humans or animals.
Parameters of water purity

Purified water is usually produced by the purification of potable water (city water) or
natural water. The impurities that may need to be removed are:

inorganic ions (typically monitored as electrical conductivity or resistivity or

specific tests)

organic compounds (typically monitored as TOC or by specific tests)

bacteria (monitored by total viable counts or epifluorescence)

endotoxins and nucleases (monitored by LAL or specific enzyme tests)

particulates (typically controlled by filtration)

gases (typically managed by degassing when required)

Raw water
Raw water is natural water found in the environment, such as rainwater, ground water,
and water from bodies like lakes and rivers. Water in this form is considered raw, as
opposed to water which has beentreated before consumption, such as drinking water or
water which has been used in an industrial process, such aswaste water. Millions of
people in developing countries rely on untreated raw water for their water supply,
sometimes purifying it by boiling.
Raw water flushing is a system for water conservation.
Composition

The composition of raw water is naturally variable but commonly contains one or more of
the following significant contaminants in the form of dissolved ions, particles and living
organisms.

Humic acid and other complex acids resulting from plant decay. These occur in
peat and soil and are significant in discolouring the water.

Minerals which make water hard. Most common are carbonates of calcium and
magnesium.

Particles of clay and silt.

Microorganisms such as bacteria, viruses, protozoa and their cysts.

Dissolved air molecules, especially oxygen

Salt, which makes water brackish, having more salinity than fresh water, but not
as much as seawater.

CHAPTER -4
MATERIALS & METHODS

MATERALS
S.NO
1
2
3

STERILIZED GLASSWARES
Glass plates
Test tubes
Measuring cylinder

4
5

Flasks
Screw cap bottles

s.no

instruments

1.

Weighing balance

2.

Autoclave

3.

Laminar air flow

4.

BOD (temp : 22 to 250C)

5.

Incubator(temp: 30 to 350C)

6.

Incubator (temp:40 to 450C)

7.

Micro pipettes

8.

Colony counter

s.no

Media

1.
2.

SOYABEAN CASEIN DIGEST MEDIUM

3.

SABOURAUD DEXTROSE AGAR

4.

MACCONKEY BROTH

SOYABEAN CASEIN DIGEST AGAR

5.

MACCONKEY AGAR

6.

MANNITOL SALT AGAR

7.

CETRIMIDE AGAR

8.

XYLOSE LYSINE DEXTROSE AGAR

9.

GN BROTH

10.

EEBM

11.

VRBGA

12.

TRIPLE SUGAR IRON AGAR

13.

PEPTONE WATER

WATER TESTING
Introduction:
Water contains many bacteria as these are found in streams, lakes and mountain
waters that also comprises of innumerable number of autotrophs and saprophytic
heterotrophs. Water becomes contaminated by intestinal pathogen such as COLI from group
of bacteria, SALMONELLA, VIBRIO and dysentery causing BACILLI.
Therefore water supply has to be checked for microbiological point of view.
In IND-SWIFT LIMITED water supply is checked regularly on alternate basis there are many
sampling points at different stages and at different locations from where the water sample is
taken for pathogen testing. First raw water and drinking water is checked. DM
(DEMINERALISED) water is used for the preparation of bulk products so that it is free from
any contamination or it itself does not add any contaminants to the products.

SAMPLING:
Sampling is done by quality control chemist and microbiologist. They are well
experienced person however, proper sampling require a most careful attention.
For sampling glass bottles are used they are of good material which is suitable for
sterilization. Sample bottles are ide mouthed and round they are covered by the caps. Bottles
shall be sterilized at 121oc for 30 min.
Before drawing a sample, wear sterile clothes, gloves and allow stream of water to flow out
for about 5 min to obtain a uniform flow. Bring the sample bottle on to ant part of outlet
sampling point. Fill the bottle to volume and cap it tightly.

BACTERIOLOGICAL TESTING:
There is number of pathogenic microorganisms responsible for contamination of water.
They are relatively easy to identify.
Mainly three tests performed to identify these. These are following-

Total aerobic bacterial cout.

Pathogen identification.
Confirmatory test.
Total aerobic bacterial count:
For water soluble products dissolve or diluted 10 ml of preparation being examined in
buffered sodium chloride peptone solution pH 7 or any suitable medium shown to have no
antimicrobial activity under the condition of test and adjust the volume to 100 ml with same
medium.