British

Pharmacopoeia Volume I & II Monographs: Medicinal and Pharmaceutical Substances Isoniazid

General Notices

(Ph. Eur. monograph 0146)

C6H7N3O

137.1

54-85-3

Action and use Antituberculosis drug. Preparations Isoniazid Injection Isoniazid Oral Solution Isoniazid Tablets Ph Eur DEFINITION Isoniazid contains not less than 99.0 per cent and not more than the equivalent of 101.0 per cent of pyridine-4-carbohydrazide, calculated with reference to the dried substance. CHARACTERS A white or almost white, crystalline powder or colourless crystals, freely soluble in water, sparingly soluble in alcohol. IDENTIFICATION First identification A, B. Second identification A, C. A. Melting point (2.2.14): 170 C to 174 C.

B. Examine by infrared absorption spectrophotometry (2.2.24), comparing with the spectrum obtained with isoniazid CRS. C. Dissolve 0.1 g in 2 mL of water R and add 10 mL of a warm 10 g/L solution of vanillin R. Allow to stand and scratch the wall of the test tube with a glass rod. A yellow precipitate is formed, which, after recrystallisation from 5 mL of alcohol (70 per cent V/V) R and drying at 100 C to 105 C, melts (2.2.14) at 226 C to 231 C. TESTS Solution S Dissolve 2.5 g in carbon dioxide-free water R and dilute to 50 mL with the same solvent. Appearance of solution Solution S is clear (2.2.1) and not more intensely coloured than reference solution BY7 (2.2.2, Method II). pH (2.2.3) The pH of solution S is 6.0 to 8.0. Hydrazine and related substances Examine by thin-layer chromatography (2.2.27), using silica gel GF254 R as the coating substance. Test solution Dissolve 1.0 g of the substance to be examined in a mixture of equal volumes of acetone R and water R and dilute to 10.0 mL with the same mixture of solvents. Reference solution Dissolve 50.0 mg of hydrazine sulfate R in 50 mL of water R and dilute to 100.0 mL with acetone R. To 10.0 mL of this solution add 0.2 mL of the test solution and dilute to 100.0 mL with a mixture of equal volumes of acetone R and water R. Apply separately to the plate 5 L of each solution and develop over a path of 15 cm using a mixture of 10 volumes of water R, 20 volumes of acetone R, 20 volumes of methanol R and 50 volumes of ethyl acetate R. Allow the plate to dry in air and examine in ultraviolet light at 254 nm. Any spot in the chromatogram obtained with the test solution, apart from the principal spot, is not more intense than the spot in the chromatogram obtained with the reference solution (0.2 per cent). Spray the plate with dimethylaminobenzaldehyde solution R1. Examine in daylight. An additional spot, corresponding to hydrazine, appears in the chromatogram obtained with the reference solution. Any corresponding spot in the chromatogram obtained with the test solution is not more intense than the spot corresponding to hydrazine in the chromatogram obtained with the reference solution (0.05 per cent). Heavy metals (2.4.8) 2.0 g complies with limit test C for heavy metals (10 ppm). Prepare the standard using 2 mL of lead standard solution (10 ppm Pb) R. Loss on drying (2.2.32)

Not more than 0.5 per cent, determined on 1.00 g by drying in an oven at 105 C. Sulfated ash (2.4.14) Not more than 0.1 per cent, determined on 1.0 g. ASSAY Dissolve 0.250 g in water R and dilute to 100.0 mL with the same solvent. To 20.0 mL of the solution add 100 mL of water R, 20 mL of hydrochloric acid R, 0.2 g of potassium bromide R and 0.05 mL of methyl red solution R. Titrate dropwise with 0.0167 M potassium bromate, shaking continuously, until the red colour disappears. 1 mL of 0.0167 M potassium bromate is equivalent to 3.429 mg of C6H7N3O.

British Pharmacopoeia Volume III Formulated Preparations: Specific Monographs

Isoniazid Tablets General Notices Action and use Antituberculosis drug. DEFINITION Isoniazid Tablets contain Isoniazid. The tablets comply with the requirements stated under Tablets and with the following requirements. Content of isoniazid, C6H7N3O 95.0 to 105.0% of the stated amount. IDENTIFICATION Shake a quantity of the powdered tablets containing 0.1 g of Isoniazid with 10 mL of ethanol (96%) for 15 minutes, centrifuge and decant the supernatant liquid. Extract the residue with two further 10-mL quantities of ethanol (96%) and evaporate the combined extracts to dryness. The infrared absorption spectrum of the residue, Appendix II A, is concordant with the reference spectrum of isoniazid (RS 196). TEST Dissolution Comply with the requirements for Monographs of the British Pharmacopoeia in the dissolution test for tablets and capsules, Appendix XII B1. TEST CONDITIONS (a) Use Apparatus 1, rotating the basket at 100 revolutions per minute. (b) Use 900 mL of water, at a temperature of 37, as the medium. PROCEDURE After 45 minutes withdraw a 10 mL sample of the medium and measure the absorbance of the filtered sample, suitably diluted if necessary, at the maximum at 263 nm, Appendix II B. DETERMINATION OF CONTENT Calculate the total content of isoniazid, C6H7N3O, in the medium taking 307 as the value of A(1%, 1 cm) at the maximum at 263 nm. ASSAY

Weigh and powder 20 tablets. Dissolve a quantity of the powder containing 150 mg of Isoniazid as completely as possible in water, filter and wash the residue with sufficient water to produce 250 mL. To 50 mL of the resulting solution add 50 mL of water, 20 mL of hydrochloric acid and 0.2 g of potassium bromide and titrate with 0.0167M potassium bromate VS determining the end point electrometrically. Each mL of 0.0167M potassium bromate VS is equivalent to 3.429 mg of C6H7N3O. STORAGE Isoniazid Tablets should be protected from light.

USP monographs Isoniazid (eye'' soe nye' a zid).

C6H7N3O

137.14

4-Pyridinecarboxylic acid, hydrazide. Isonicotinic acid hydrazide [54-85-3]. Isoniazid contains not less than 98.0 percent and not more than 102.0 percent of C6H7N3O, calculated on the dried basis. Packaging and storagePreserve in tight, light-resistant containers. Store at 25 , excursions permitted between 15 and 30 . USP REFERENCE STANDARDS 11

USP Isoniazid RS Identification A: Infrared Absorption 197K .

B: Transfer about 50 mg of it to a 500-mL volumetric flask, add water to volume, and mix. Transfer 10.0 mL of the resulting solution to a 100-mL volumetric flask, add 2.0 mL of 0.1 N hydrochloric acid, dilute with water to volume, and mix to obtain a 1 in 100,000 solution: the UV absorption spectrum of the solution so obtained exhibits maxima and minima only at the same wavelengths as that of a similar solution of USP Isoniazid RS, concomitantly measured. MELTING RANGE
PH

741 : between 170 and 173 .

791 : between 6.0 and 7.5, in a solution (1 in 10). 731 Dry it at 105 for 4 hours: it loses not more than 1.0% of its weight. 281 : not more than 0.2%. 231 : 0.002%.

LOSS ON DRYING

RESIDUE ON IGNITION

HEAVY METALS, Method II Assay

Mobile phaseDissolve 4.4 g of docusate sodium in 600 mL of methanol, add 400 mL of water, adjust with 2 N sulfuric acid to a pH of 2.5, and mix. Make adjustments if necessary (see System Suitability under Chromatography 621 ).

Standard preparationDissolve an accurately weighed quantity of USP Isoniazid RS in Mobile phase, and quantitatively dilute with Mobile phase to obtain a solution having a known concentration of about 0.32 mg per mL. Assay preparationTransfer about 16 mg of Isoniazid, accurately weighed, to a 50-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix. Chromatographic system (see CHROMATOGRAPHY 621 )The liquid chromatograph is equipped

with a 254-nm detector and a 4.6-mm 25-cm column that contains packing L1. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the column efficiency determined from the isoniazid peak is not less than 1800 theoretical plates; the tailing factor for the isoniazid peak is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%. ProcedureSeparately inject equal volumes (about 10 L) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the quantity, in mg, of C6H7N3O in the portion of Isoniazid taken by the formula: 50C(rU / rS) in which C is the concentration, in mg per mL, of USP Isoniazid RS in the Standard preparation; and rU and rS are the peak responses of isoniazid obtained from the Assay preparation and the Standard preparation, respectively. Auxiliary Information Please check for your question in the FAQs before contacting USP. Topic/Question Monograph Contact Expert Committee Behnam Davani, Ph.D., M.B.A. (SM12010) Monographs - Small Molecules 1

Senior Scientific Liaison 1-301-816-8394 Reference Standards RS Technical Services 1-301-816-8129 rstech@usp.org USP34NF29 Page 3203 Pharmacopeial Forum: Volume No. 29(6) Page 1912 Chromatographic Column ISONIAZID Chromatographic columns text is not derived from, and not part of, USP 34 or NF 29.

Isoniazid Tablets Isoniazid Tablets contain not less than 90.0 percent and not more than 110.0 percent of the labeled amount of isoniazid (C6H7N3O). Packaging and storagePreserve in well-closed, light-resistant containers. USP REFERENCE STANDARDS 11

USP Isoniazid RS Identification A: The retention time of the isoniazid peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay. B: Transfer a portion of finely powdered Tablets, equivalent to about 50 mg of isoniazid, to a 500mL volumetric flask. Add water to volume, mix, and filter a portion of the mixture. Proceed as directed in Identification test B under Isoniazid, beginning with Transfer 10.0 mL of the resulting solution to a 100-mL volumetric flask. DISSOLUTION 711

Medium: 0.01 N hydrochloric acid; 900 mL. Apparatus 1: 100 rpm. Time: 45 minutes. ProcedureDetermine the amount of C6H7N3O dissolved by employing UV absorption at the wavelength of maximum absorbance at about 263 nm on filtered portions of the solution under test, suitably diluted with Dissolution Medium, in comparison with a Standard solution having a known concentration of USP Isoniazid RS in the same Medium. TolerancesNot less than 80% (Q) of the labeled amount of C6H7N3O is dissolved in 45 minutes. UNIFORMITY OF DOSAGE UNITS 905 : meet the requirements.

Procedure for content uniformityTransfer 1 finely powdered Tablet to a 500-mL volumetric flask with the aid of 200 mL of water. Shake by mechanical means for 30 minutes, add water to volume, and mix. Filter, and discard the first 20 mL of the filtrate. Dilute a portion of the filtrate quantitatively and stepwise, if necessary, with a 3 in 100 mixture of 0.1 N hydrochloric acid and water to obtain a solution containing about 10 g per mL. Dissolve an accurately weighed quantity of USP Isoniazid RS in a volume of water corresponding to that used to dissolve a similar amount of isoniazid from the Tablet, and dilute quantitatively and stepwise, if necessary, with a 3 in 100 mixture of 0.1 N hydrochloric acid and water to obtain a Standard solution having a known concentration of about 10 g per mL. Concomitantly determine the absorbances of both solutions in 1-cm cells at the wavelength of maximum absorbance at about 263 nm, with a suitable spectrophotometer, using water as the blank. Calculate the quantity, in mg, of isoniazid (C6H7N3O) in the Tablet taken by the formula:

(TC / D)(AU / AS) in which T is the labeled quantity, in mg, of isoniazid in the Tablet; C is the concentration, in g per mL, of USP Isoniazid RS in the Standard solution; D is the concentration, in g per mL, of isoniazid in the solution from the Tablet, based on the labeled quantity per Tablet and the extent of dilution; and AU and AS are the absorbances of the solution from the Tablet and the Standard solution, respectively. Assay Buffer solutionPrepare a 0.1 M monobasic potassium phosphate solution, adjust with 10 N sodium hydroxide to a pH of 6.9, add sufficient triethanolamine to obtain a solution having a known concentration of 0.2 mM of triethanolamine, and mix. Mobile phasePrepare a filtered and degassed mixture of Buffer solution and methanol (95:5). Make adjustments if necessary (see System Suitability under Chromatography 621 ).

Standard preparationDissolve an accurately weighed quantity of USP Isoniazid RS in Mobile phase, and dilute quantitatively, and stepwise, if necessary, with Mobile phase to obtain a solution having a known concentration of about 0.32 mg per mL. Assay preparationWeigh and finely powder not fewer than 20 Tablets. Transfer an accurately weighed portion of the powder, equivalent to 32 mg of isoniazid, to a 100-mL volumetric flask, add 40 mL of Mobile phase, and sonicate for 10 minutes. Cool to room temperature, dilute with Mobile phase to volume, and centrifuge for 5 minutes. Chromatographic system (see Chromatography 621 )The liquid chromatograph is equipped

with a 254-nm detector and a 3.9-mm 30-cm column that contains packing L1. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the capacity factor, k, is not less than 2.35; the column efficiency is not less than 1800 theoretical plates; the tailing factor is not more than 1.5; and the relative standard deviation for replicate injections is not more than 1.0%. ProcedureSeparately inject equal volumes (about 20 L) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the quantity, in mg, of isoniazid (C6H7N3O) in the portion of Tablets taken by the formula: 100C(rU / rS) in which C is the concentration, in mg per mL, of USP Isoniazid RS in the Standard preparation; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively. Auxiliary Information Please check for your question in the FAQs before contacting USP. Topic/Question Monograph Contact Behnam Davani, Ph.D., M.B.A. Expert Committee (SM12010) Monographs - Small Molecules 1

Topic/Question

Contact Senior Scientific Liaison 1-301-816-8394 Reference RS Technical Services Standards 1-301-816-8129 rstech@usp.org Margareth R.C. Marques, 711 Ph.D. Senior Scientific Liaison 1-301-816-8106 USP34NF29 Page 3204 Chromatographic Column ISONIAZID TABLETS

Expert Committee

(GCDF2010) General Chapters - Dosage Forms

Chromatographic columns text is not derived from, and not part of, USP 34 or NF 29.