JP XVI

General Tests / 1.07

Heavy Metals Limit Test

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pull out C carefully, add 2 drops of hydrazine hydrate to the test solution, put C on A, and decolorize the solution by vigorous shaking. Transfer the content of A to a beaker, wash C, B and the inner side of A with 25 mL of 2-propanol, and transfer the washings to the above beaker. To this solution add 1 drop of bromophenol blue TS, then add dilute nitric acid dropwise until a yellow color develops, and titrate <2.50> with 0.005 mol/L silver nitrate VS according to the potentiometric tiration. Perform the test with the blank solution in the same manner, and make any necessary correction. Each mL of 0.005 mol/L silver nitrate VS 0.6345 mg of I 3.3. Fluorine Apply a small amount of water to the upper part of A, pull out C carefully, transfer the test solution and the blank solution to 50 mL volumetric flasks separately, wash C, B and the inner side of A with water, add the washings and water to make 50 mL, and use these solutions as the test solution and the correction solution. Pipet the test solution (V mL) equivalent to about 30 mg of fluorine, V mL of the correction solution and 5 mL of standard fluorine solution, transfer to 50-mL volumetric flasks separately, add 30 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution, pH 4.3 and cerium (III) nitrate TS (1:1:1), add water to make 50 mL, and allow to stand for 1 hour. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a blank prepared with 5 mL of water in the same manner. Determine the absorbances, AT, AC and AS, of the subsequent solutions of the test solution, the correction solution and the standard solution at 600 nm. Fig. 1.06-1 Amount (mg) of fluorine (F) in the test solution amount (mg) of fluorine in 5 mL of the standard solution the white smoke in A vanishes completely, allow to stand for 15 to 30 minutes, and designate the resulting solution as the test solution. Prepare the blank solution in the same manner, without sample. Procedure of determination Unless otherwise specified in the monograph, perform the test as follows. 3.1. Chlorine and bromine Apply a small amount of water to the upper part of A, pull out C carefully, and transfer the test solution to a beaker. Wash C, B and the inner side of A with 15 mL of 2propanol, and combine the washings with the test solution. To this solution add 1 drop of bromophenol blue TS, add dilute nitric acid dropwise until a yellow color develops, then add 25 mL of 2-propanol, and titrate <2.50> with 0.005 mol/L silver nitrate VS according to the potentiometric titration. Perform the test with the blank solution in the same manner, and make any necessary correction. Each mL of 0.005 mol/L silver nitrate VS 0.1773 mg of Cl Each mL of 0.005 mol/L silver nitrate VS 0.3995 mg of Br 3.2. Iodine Apply a small amount of water to the upper part of A, 3.

AT AC 50 AS V

Standard Fluorine Solution: Dry sodium fluoride (standard reagent) in a platinum crucible between 5009 and C C 5509 for 1 hour, cool it in a desiccator (silica gel), weigh accuraly about 66.3 mg of it, and dissolve in water to make exactly 500 mL. Pipet 10 mL of this solution, and dilute with sufficient water to make exactly 100 mL. 3.4. Sulfur Apply a small amount of water to the upper part of A, pull out C carefully, and wash C, B and the inner side of A with 15 mL of methanol. To this solution add 40 mL of methanol, then add exactly 25 mL of 0.005 mol/L barium perchlorate VS, allow to stand for 10 minutes, add 0.15 mL of arsenazo III TS with a measuring pipet, and titrate <2.50> with 0.005 mol/L sulfuric acid VS. Perfrom the test with the blank solution in the same manner. Each mL of 0.005 mol/L barium perchlorate VS 0.1604 mg of S

1.07 Heavy Metals Limit Test

Heavy Metals Limit Test is a limit test of the quantity of heavy metals contained as impurities in drugs. The heavy metals are the metallic inclusions that are darkened with so-

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1.08

Nitrogen Determination (Semimicro-Kjeldahl Method) / General Tests

JP XVI

dium sulfide TS in acidic solution, as their quantity is expressed in terms of the quantity of lead (Pb). In each monograph, the permissible limit for heavy metals (as Pb) is described in terms of ppm in parentheses. Preparation of test solutions and control solutions Unless otherwise specified, test solutions and control solutions are prepared as directed in the following: 1.1. Method 1 Place an amount of the sample, directed in the monograph, in a Nessler tube. Dissolve in water to make 40 mL. Add 2 mL of dilute acetic acid and water to make 50 mL, and designate it as the test solution. The control solution is prepared by placing the volume of Standard Lead Solution directed in the monograph in a Nessler tube, and adding 2 mL of dilute acetic acid and water to make 50 mL. 1.2. Method 2 Place an amount of the sample, directed in the monograph, in a quartz or porcelain crucible, cover loosely with a lid, and carbonize by gentle ignition. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid, heat cautiously until white fumes are no longer evolved, and incinerate by ignition between 5009 and 6009 Cool, add 2 mL C C. of hydrochloric acid, evaporate to dryness on a water bath, moisten the residue with 3 drops of hydrochloric acid, add 10 mL of hot water, and warm for 2 minutes. Then add 1 drop of phenolphthalein TS, add ammonia TS dropwise until the solution develops a pale red color, add 2 mL of dilute acetic acid, filter if necessary, and wash with 10 mL of water. Transfer the filtrate and washings to a Nessler tube, and add water to make 50 mL. Designate it as the test solution. The control solution is prepared as follows: Evaporate a mixture of 2 mL of nitric acid, 5 drops of sulfuric acid and 2 mL of hydrochloric acid on a water bath, further evaporate to dryness on a sand bath, and moisten the residue with 3 drops of hydrochloric acid. Hereinafter, proceed as directed in the test solution, then add the volume of Standard Lead Solution directed in the monograph and water to make 50 mL. 1.3. Method 3 Place an amount of the sample, directed in the monograph, in a quartz or porcelain crucible, heat cautiously, gently at first, and then incinerate by ignition between 5009 C C. and 6009 After cooling, add 1 mL of aqua regia, evaporate to dryness on a water bath, moisten the residue with 3 drops of hydrochloric acid, add 10 mL of hot water, and warm for 2 minutes. Add 1 drop of phenolphthalein TS, add ammonia TS dropwise until the solution develops a pale red color, add 2 mL of dilute acetic acid, filter if necessary, wash with 10 mL of water, transfer the filtrate and washings to a Nessler tube, and add water to make 50 mL. Designate it as the test solution. The control solution is prepared as follows: Evaporate 1 mL of aqua regia to dryness on a water bath. Hereinafter, proceed as directed for the test solution, and add the volume of Standard Lead Solution directed in the monograph and water to make 50 mL. 1.4. Method 4 Place an amount of the sample, directed in the monograph, in a platinum or porcelain crucible, mix with 10 mL of a solution of magnesium nitrate hexahydrate in ethanol 1.

(95) (1 in 10), fire the ethanol to burn, and carbonize by gradual heating. Cool, add 1 mL of sulfuric acid, heat carefully, and incinerate by ignition between 5009 and 6009 C C. If a carbonized substance remains, moisten with a small amount of sulfuric acid, and incinerate by ignition. Cool, dissolve the residue in 3 mL of hydrochloric acid, evaporate on a water bath to dryness, wet the residue with 3 drops of hydrochloric acid, add 10 mL of water, and dissolve by warming. Add 1 drop of phenolphthalein TS, add ammonia TS dropwise until a pale red color develops, then add 2 mL of dilute acetic acid, filter if necessary, wash with 10 mL of water, transfer the filtrate and the washing to a Nessler tube, add water to make 50 mL, and use this solution as the test solution. The control solution is prepared as follows: Take 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and fire the ethanol to burn. Cool, add 1 mL of sulfuric acid, heat carefully, and ignite between 5009 C C. and 6009 Cool, and add 3 mL of hydrochloric acid. Hereinafter, proceed as directed in the test solution, then add the volume of Standard Lead Solution directed in the monograph and water to make 50 mL. 2. Procedure Add 1 drop of sodium sulfide TS to each of the test solution and the control solution, mix thoroughly, and allow to stand for 5 minutes. Then compare the colors of both solutions by viewing the tubes downward or transversely against a white background. The test solution has no more color than the control solution.

1.08 Nitrogen Determination (Semimicro-Kjeldahl Method)

Nitrogen Determination is a method to determine nitrogen in an organic substance in which the nitrogen is converted into ammonia nitrogen by thermal decomposition of the organic substance with sulfuric acid, and the ammonia liberated by alkali and trapped by distillation with steam is determined by titration. 1. Apparatus Use the apparatus illustrated in Fig. 1.08-1. It is thoroughly constructed of hard glass, and ground glass surfaces may be used for joints. All rubber parts used in the apparatus should be boiled for 10 to 30 minutes in sodium hydroxide TS and for 30 to 60 minutes in water, and finally washed thoroughly with water before use. Alternatively, apparatus can be used in which some of the procedures, such as digestion of organic substances, distillation of the liberated ammonia, and endpoint detection methods in titrimetry (e.g., potentiometric titration or titration by colorimeter) are automated. 2. System suitability If an automated apparatus is used, it is necessary to confirm periodically the suitability of the apparatus according to the following method: Weigh accurately about 1.7 g of amidosulfuric acid (standard reagent), previously dried in a desiccator (in vacuum, silica gel) for about 48 hours, dissolve in water to make exactly 200 mL. Pipet 2 mL of this solution, and transfer to a