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.

Methyl Red Solution   Dissolve 50 mg of methyl red in a mixture of 1.86 ml of 0.1M sodium
hydroxide and 50 ml of ethanol (96%) and add sufficient water to produce 100 ml.
Complies with the following test.

Sensitivity  A mixture of 0.1 ml of the solution, 100 ml of carbon dioxide-free water and 0.05 ml of
0.02M hydrochloric acid VS is red. Not more than 0.1 ml of 0.02M sodium hydroxide VS is required to
change the colour of the solution to yellow.

Colour change pH 4.4 (red) to pH 6.0 (yellow).

Hydrochloric Acid VS

Hydrochloric Acid VS   HCl = 36.46

For a 0.1M solution  Dilute 8.5 ml of hydrochloric acid with sufficient water to produce 1000 ml.
Ascertain its exact concentration in the following manner. Dissolve 0.1 g of anhydrous sodium
carbonate in 20 ml of water , add 0.1 ml of methyl orange solution and titrate with the hydrochloric
acid until the solution becomes reddish yellow. Boil for 2 minutes and continue the titration until the
reddish yellow colour is restored. Each ml of 0.1M hydrochloric acid VS is equivalent to 5.30 mg of
Na2CO3.

For a 1M solution  Dilute 103.0 g of hydrochloric acid to 1000 ml with water .

Ascertain its exact concentration as described for the 0.1M solution using 1.000 g of anhydrous
sodium carbonate dissolved in 50 ml of water . Each ml of 1M hydrochloric acid VS is equivalent to
53.00 mg of Na2CO3.

For a 2M solution  Dilute 206.0 g of hydrochloric acid to 1000 ml with water .

For a 3M solution  Dilute 309.0 g of hydrochloric acid to 1000 ml with water .

For a 6M solution  Dilute 618.0 g of hydrochloric acid to 1000 ml with water .

Sodium Hydroxide VS

Sodium Hydroxide VS   NaOH = 40.00

For a 0.1M solution  Dissolve 4.2 g of sodium hydroxide in sufficient carbon dioxide-free water to
produce 1000 ml.
Ascertain its exact concentration immediately before use in the following manner. Titrate 20 ml of the
solution with 0.1M hydrochloric acid VS using the indicator prescribed in the assay in which the
solution is to be used. Each ml of 0.1M hydrochloric acid VS is equivalent to 4.00 mg of NaOH.
When a carbonate-free solution is specified it is prepared using the following method. Dissolve
sodium hydroxide in an equal weight of water and allow to stand overnight. Taking precautions to
avoid absorption of carbon dioxide, siphon off or decant the clear supernatant liquid and dilute with
carbon dioxide-free water to the desired molarity. The solution complies with the following test.
 Titrate 20 ml of hydrochloric acid VS of the same molarity as the solution being examined with the
sodium hydroxide solution using 0.5 ml of phenolphthalein solution as indicator. At the end point add
just sufficient of the acid to discharge the pink colour and boil to reduce the volume to 20 ml. Add,
whilst boiling, sufficient of the acid again to discharge the pink colour which does not reappear after
prolonged boiling. Not more than 0.1 ml of the acid is required.

For a 1M solution  Dissolve 42 g of sodium hydroxide in sufficient carbon dioxide-free water to

produce 1000 ml.
Ascertain its exact concentration immediately before use in the following manner. Titrate 20 ml of the
solution with 1M hydrochloric acid VS using the indicator prescribed in the assay in which the
solution is to be used. Each ml of 1M hydrochloric acid VS is equivalent to 40.00 mg of NaOH.
When 0.1M sodium hydroxide VS is used in the assay of halide salts of organic bases, ascertain its
exact concentration in the following manner. Dissolve 0.100 g of benzoic acid in a mixture of 5 ml of
0.01M hydrochloric acid and 50 ml of ethanol (96%). Titrate with the solution being examined and
note the volume added between the two points of inflection. Each ml of 0.1M sodium hydroxide VS is
equivalent to 12.21 mg of C7H6O.

Hydrochloric Acid   HCl = 36.46 (7647-01-0)

Where no molarity is indicated use analytical reagent grade of commerce with a relative density of
about 1.18, containing not less than 35% w/w and not more than 38% w/w of HCl and about 11.5M in
strength.
A colourless, fuming liquid.
Solutions of molarity xM should be prepared by diluting 85x ml of hydrochloric acid to 1000 ml with
water .
Store in a container of polyethylene or other non-reacting material at a temperature not exceeding
30°.

Potassium Bromate

Potassium Bromate   KBrO3 = 167.0 (7758-01-2)

Analytical reagent grade of commerce.
White, granular powder or crystals.

Potassium Bromide

Potassium Bromide   KBr = 119.0 (7758-02-3)

Analytical reagent grade of commerce.
Potassium bromide used for infrared absorption spectrophotometry complies with the following
requirement.
The infrared absorption spectrum, Appendix II A, of a disc 2 mm thick prepared from material
previously dried at 250° for 1 hour has a substantially flat baseline over the range 4000 to 620 cm–1;
it exhibits no maxima with an absorbance greater than 0.02 above the baseline with the exception of
maxima due to water at 3440 and 1630 cm–1.
Merck index
Absorption maximum: uv max (water): 266 nm (E1%1cm 378); (0.01N HCl): 265 nm (E1%1cm ~420)
Toxicity data: LD50 in mice (mg/kg): 151 i.p., 149 i.v. (Jenney, Pfeiffer)

Derivative Type: 4-Aminosalicylate

CAS Registry Number: 2066-89-9
Additional Names: Pasiniazide
Manufacturers' Codes: GEWO-399
Trademarks: Paraniazide (Hepatrol); Dipasic (Gewo)
Molecular Formula: C13H14N4O4
Molecular Weight: 290.27.
Percent Composition: C 53.79%, H 4.86%, N 19.30%, O 22.05%
Literature References: An equimolecular salt produced by dissolving the components in hot dil alc,
followed by cooling and evapn: Charonnat, Boime, Compt. Rend. 236, 2140 (1953). Prepn: CH
303085 (1955 to Hoffmann-La Roche).
Properties: Yellow crystals from methanol or ethanol, mp 142-144°. Sparingly sol in water. uv max
272, 303 nm (E1%1cm 550, 445).
Melting point: mp 142-144°
Absorption maximum: uv max 272, 303 nm (E1%1cm 550, 445)

Derivative Type: Methanesulfonate

CAS Registry Number: 13447-95-5
CAS Name: 4-Pyridinecarboxylic acid 2-(sulfomethyl)hydrazide
Additional Names: methaniazide
Molecular Formula: C7H9N3O4S
Molecular Weight: 231.23.
Percent Composition: C 36.36%, H 3.92%, N 18.17%, O 27.68%, S 13.87%
Literature References: Prepn: Logemann, US 2759944 (1956 to Carlo Erba).
Properties: Crystals, dec 187-189°.

Derivative Type: Methanesulfonate sodium

CAS Registry Number: 3804-89-5
Additional Names: Sodium isonicotinylhydrazide methanesulfonate
Trademarks: Neoiscotin (Daiichi)
Molecular Formula: C7H8N3NaO4S
Molecular Weight: 253.21.
Percent Composition: C 33.20%, H 3.18%, N 16.60%, Na 9.08%, O 25.27%, S 12.66%
Properties: Yellow crystals from water, dec 164-167°.

Derivative Type: Methanesulfonate calcium

CAS Registry Number: 6059-26-3
Trademarks: Neo-Tizide (Farmitalia Carlo Erba)
Molecular Formula: C14H16CaN6O8S2
Molecular Weight: 500.52.
Percent Composition: C 33.60%, H 3.22%, Ca 8.01%, N 16.79%, O 25.57%, S 12.81%
Properties: Crystals, dec 215-220°.

Therap-Cat: Antibacterial (tuberculostatic).

Therap-Cat-Vet: Antibacterial (tuberculostatic); anti-actinomycotic agent.