色氨酸欧洲药典EP8.0.pdfVIP

Tryptophan EUROPEAN PHARMACOPOEIA 8.0 Reference solution. Dissolve 25.0 mg of trypsin BRP in 0.001 M IDENTIFICATION hydrochloric acid and dilute to 25.0 mL with the same acid. First identification: A, B. Store the solutions at 0-5 °C. Warm 1 mL of each solution to Second identification: A, C, D. about 25 °C over 15 min and use 50 μL of each solution for A. Specific optical rotation (see Tests). each titration. Carry out the titration in an atmosphere of nitrogen. Transfer 10.0 mL of 0.0015 M borate buffer solution B. Infrared absorption spectrophotometry (2.2.24). pH 8.0 R to the reaction vessel and, while stirring, add 1.0 mL Preparation : discs. of a freshly prepared 6.86 g/L solution of benzoylarginine Comparison : tryptophan CRS. ethyl ester hydrochloride R. When the temperature is steady C. Examine the chromatograms obtained in the test for at 25.0 ± 0.1 °C (after about 5 min) adjust to pH 8.0 exactly ninhydrin-positive substances. with 0.1 M sodium hydroxide. Add 50 μL of the test solution Results : the principal spot in the chromatogram obtained and start a timer. Maintain at pH 8.0 by the addition of 0.1 M with test solution (b) is similar in position, colour and size sodium hydroxide, the tip of the microburette being immersed to the principal spot in the chromatogram obtained with in the solution; note the volume added every 30 s. Follow the reference solution (a). reaction for 8 min. Calculate the volume of 0.1 M sodium hydroxide used per second. Carry out a titration in the same D. Dissolve about 20 mg in 10 mL of water R. Add 5 mL manner using the reference solution and calculate the volume