789789JP XVOcial Monographs/ Kainic Acid Hydrate1/15 mol/L phosphate buer solution, pH 5.6 to make ex- actly 50 mL, and use this solution as the standard stock solu- tion. Keep the standard stock solution at 59C or below, and use within 3 days. Take exactly a suitable amount of the stan- dard stock solution before use, add 1/15 mol/L phosphate buer solution, pH 5.6 to make solutions so that each mL contains 80mg (potency) and 20mg (potency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. (iv)Sample solutionsWeigh accurately an amount of Josamycin Propionate, equivalent to about 20 mg (potency), dissolve in 10 mL of methanol, add 1/15 mol/L phosphate buer solution, pH 5.6 to make exactly 50 mL. Take exactly a suitable amount of this solution, add 1/15 mol/L phos- phate buer solution, pH 5.6 to make solutions so that each mL contains 80mg (potency) and 20mg (potency), and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.Containers and storageContainersTight containers. StorageLight-resistant.Kainic Acid Hydrate?C10H15NO4.H2O: 231.25 (2S,3S,4S)-3-(Carboxymethyl)- 4-(1-methylethenyl)pyrrolidine-2-carboxylic acid monohydrate487-79-6, anhydrideKainic Acid Hydrate, when dried, contains not less than 99.0zof kainic acid (C10H15NO4: 213.23).DescriptionKainic Acid Hydrate occurs as white crystals or crystalline powder. It is odorless, and has an acid taste. It is sparingly soluble in water and in warm water, very slightly soluble in acetic acid (100) and in ethanol (95), and practically insoluble in diethyl ether. It dissolves in dilute hydrochloric acid and in sodium hydroxide TS. The pH of its solution (1 in 100) is between 2.8 and 3.5. Melting point: about 2529C (with decomposition).Identication(1)To 5 mL of a solution of Kainic Acid Hydrate (1 in 5000) add 1 mL of ninhydrin TS, and warm in a water bath at a temperature between 609C and 709C for 5 minutes: a yellow color is produced. (2)Dissolve 0.05 g of Kainic Acid Hydrate in 5 mL of a- cetic acid (100), and add 0.5 mL of bromine TS: the color of bromine disappears immediately.Optical rotationa20D:?13 ?179(0.5 g, water, 50 mL, 200 mm).Purity(1)Clarity and color of solutionDissolve 0.10 g of Kainic Acid Hydrate in 10 mL of water: the solution is clear and colorless. (2)ChlorideTake 0.5 g of Kainic Acid Hydratein a platinum crucible, dissolve in 5 mL of sodium carbonate TS, and evaporate on a water bath to dryness. Heat the cruci- ble slowly at rst, and then ignite until the sample is almost incinerated. After cooling, add 12 mL of dilute nitric acid to the residue, dissolve by warming, and lter. Wash the residue with 15 mL of water, combine the washings and the ltrate, and add water to make 50 mL. Perform the test using this so- lution as the test solution. Control solution: Add 5 mL of sodium carbonate TS to 0.30 mL of 0.01 molWL hydrochloric acid VS, and proceed as directed above (not more than 0.021z). (3)SulfateDissolve 0.5 g of Kainic Acid Hydrate in 40 mL of water by warming. Cool, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con- trol solution with 0.30 mL of 0.005 molWL sulfuric acid VS (not more than 0.028z). (4)AmmoniumTake 0.25 g of Kainic Acid Hydrate, and perform the test. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02 z). (5)Heavy metalsProceed with 1.0 g of Kainic Acid Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (6)ArsenicDissolve 1.0 g of Kainic Acid Hy- drate in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 2 ppm). (7)Amino acid and other imino acidDissolve 0.10 g of Kainic Acid Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 2 mL of this solution, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test as directed under Thin- layer Chromatographywith these solutions. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with the supernatant liquid of a mixture of water, 1- butanol and acetic acid (100) (5:4:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of nin- hydrin in acetone (1 in 50) on the plate, and dry the plate at 809C for 5 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution.Loss on drying6.5 8.5z(1 g, 1059C, 4 hours).Residue on ignitionNot more than 0.1z(0.5 g).AssayWeigh accurately about 0.4 g of Kainic Acid Hy- drate, previously dried,