EUROPEAN PHARMACOPOEIA 5.

Ammonium glycyrrhizate

01/2005:1772 Reference solution (a). Dilute 1.0 ml of the test solution to 20.0 ml with the mobile phase.

AMMONIUM GLYCYRRHIZATE Ammonii glycyrrhizas

Reference solution (b). Dissolve 50 mg of ammonium glycyrrhizate CRS in the mobile phase and dilute to 50.0 ml with the mobile phase. Dilute 1.0 ml of the solution to 20.0 ml with the mobile phase. Column : size : l = 0.25 m, = 4.0 mm, stationary phase : octadecylsilyl silica gel for chromatography R (5-10 m). Mobile phase : glacial acetic acid R, acetonitrile R, water R (6:380:614 V/V/V). Flow rate : 1.2 ml/min. Detection : spectrophotometer at 254 nm. Injection : 10 l. Run time : 3 times the retention time of 18-glycyrrhizic acid.

C42H65NO16 DEFINITION

Relative retention with reference to 18-glycyrrhizic acid (retention time = about 8 min) : impurity A = about 0.8 ; Mr 840 18-glycyrrhizic acid = about 1.2. System suitability : reference solution (b) : resolution : minimum 2.0 between the peaks due to 18-glycyrrhizic acid and 18-glycyrrhizic acid. Limits : 18-glycyrrhizic acid : not more than twice the sum of the areas of the peaks in the chromatogram obtained with reference solution (a) (10.0 per cent), impurity A : not more than the sum of the areas of the peaks in the chromatogram obtained with reference solution (a) (5.0 per cent),

Mixture of ammonium 18- and 18-glycyrrhizate (ammonium salt of (20)-3-[[2-O-(-D-glucopyranosyluronic acid)--D-glucopyranosyluronic acid]oxy]-11-oxoolean-12-en29-oic acid), the 18-isomer being the main component. Content : 98.0 per cent to 102.0 per cent (anhydrous substance). CHARACTERS Appearance : white or yellowish-white, hygroscopic powder.

Solubility : slightly soluble in water, very slightly soluble any other impurity : for each impurity, not more than in anhydrous ethanol, practically insoluble in acetone. It 0.4 times the sum of the areas of the peaks in the dissolves in dilute solutions of acids and of alkali hydroxides. chromatogram obtained with reference solution (a) (2.0 per cent), IDENTIFICATION total of other impurities : not more than 1.4 times the A. Infrared absorption spectrophotometry (2.2.24). sum of the areas of the peaks in the chromatogram obtained with reference solution (a) (7.0 per cent), Comparison : ammonium glycyrrhizate CRS. B. Dissolve 0.1 g in 20 ml of water R, add 2 ml of dilute sodium hydroxide solution R and heat cautiously. On heating, the solution gives off vapours that may be identified by the alkaline reaction of wet litmus paper (2.3.1). TESTS Appearance of solution. The solution is clear (2.2.1) and not more intensely coloured than reference solution BY7 (2.2.2, Method I). disregard limit : 0.04 times the sum of the areas of the peaks in the chromatogram obtained with reference solution (a) (0.2 per cent). Heavy metals (2.4.8) : maximum 20 ppm. 1.0 g complies with limit test C. Prepare the reference solution using 2 ml of lead standard solution (10 ppm Pb) R. Water (2.5.12) : maximum 6.0 per cent, determined on 0.250 g. Sulphated ash (2.4.14) : maximum 0.2 per cent, determined on 1.0 g.

Dissolve 1.0 g in ethanol (20 per cent V/V) R and dilute to ASSAY 100.0 ml with the same solvent. Specific optical rotation (2.2.7) : + 49.0 to + 54.0 (anhydrous Dissolve 0.600 g in 60 ml of acetic acid R heating at 80 C if necessary. Cool. Titrate with 0.1 M perchloric acid, substance). determining the end-point potentiometrically (2.2.20). Dissolve 0.5 g in ethanol (50 per cent V/V) R and dilute to 1 ml of 0.1 M perchloric acid is equivalent to 84.0 mg 50.0 ml with the same solvent. of C42H65NO16. Related substances. Liquid chromatography (2.2.29). Test solution. Dissolve 0.100 g of the substance to be examined in the mobile phase and dilute to 100.0 ml with the mobile phase. General Notices (1) apply to all monographs and other texts STORAGE In an airtight container. 987

Ammonium hydrogen carbonate

EUROPEAN PHARMACOPOEIA 5.0

IMPURITIES

ASSAY Dissolve cautiously 1.0 g in 20.0 ml of 0.5 M sulphuric acid and dilute to 50 ml with water R. Boil, cool and titrate the excess of acid with 1 M sodium hydroxide, using 0.1 ml of methyl red solution R as indicator. 1 ml of 0.5 M sulphuric acid is equivalent to 79.1 mg of NH4HCO3. STORAGE Store in an airtight container. 01/2005:0594

AMOBARBITAL
A. (4,20)-3-[[2-O-(-D-glucopyranosyluronic acid)--D-glucopyranosyluronic acid]oxy]-23-hydroxy11-oxoolean-12-en-29-oic acid (24-hydroxyglycyrrhizinic acid). 01/2005:1390

Amobarbitalum

AMMONIUM HYDROGEN CARBONATE Ammonii hydrogenocarbonas

C11H18N2O3

Mr 226.3

DEFINITION Amobarbital contains not less than 99.0 per cent and NH4HCO3 Mr 79.1 not more than the equivalent of 101.0 per cent of 5-ethyl-5-(3-methylbutyl)pyrimidin-2,4,6(1H,3H,5H)-trione, DEFINITION calculated with reference to the dried substance. Ammonium hydrogen carbonate contains not less than 98.0 per cent and not more than 101.0 per cent of the CHARACTERS equivalent of ammonium hydrogen carbonate. A white, crystalline powder, very slightly soluble in water, freely soluble in alcohol, soluble in methylene chloride. It CHARACTERS forms water-soluble compounds with alkali hydroxides and A fine, white, crystalline powder or white crystals, slightly carbonates and with ammonia. hygroscopic, freely soluble in water, practically insoluble in alcohol. IDENTIFICATION It volatilises rapidly at 60 C. The volatilisation takes First identification : A, B. place slowly at ambient temperatures if the substance is Second identification : A, C, D. slightly moist. It is in a state of equilibrium with ammonium A. Determine the melting point (2.2.14) of the substance carbamate. to be examined. Mix equal parts of the substance to be examined and amobarbital CRS and determine the IDENTIFICATION melting point of the mixture. The difference between the A. It gives the reaction of carbonates and bicarbonates melting points (which are about 157 C) is not greater (2.3.1). than 2 C. B. Dissolve 50 mg in 2 ml of water R. The solution gives the B. Examine by infrared absorption spectrophotometry reaction of ammonium salts (2.3.1). (2.2.24), comparing with the spectrum obtained with amobarbital CRS. TESTS C. Examine by thin-layer chromatography (2.2.27), using Solution S. Dissolve 14.0 g in 100 ml of distilled water R. silica gel GF254 R as the coating substance. Boil to remove the ammonia, allow to cool and dilute to Test solution. Dissolve 0.1 g of the substance to be 100.0 ml with distilled water R. examined in alcohol R and dilute to 100 ml with the same Chlorides (2.4.4). Dilute 5 ml of solution S to 15 ml with solvent. water R. The solution complies with the limit test for Reference solution. Dissolve 0.1 g of amobarbital CRS in chlorides (70 ppm). alcohol R and dilute to 100 ml with the same solvent. Sulphates (2.4.13). 15 ml of solution S complies with the Apply separately to the plate 10 l of each solution. limit test for sulphates (70 ppm). Develop over a path of 18 cm using the lower layer from Iron (2.4.9). Dilute 1.8 ml of solution S to 10 ml with a mixture of 5 volumes of concentrated ammonia R, water R. The solution complies with the limit test for iron 15 volumes of alcohol R and 80 volumes of chloroform R. (40 ppm). Examine immediately in ultraviolet light at 254 nm. The Heavy metals (2.4.8). Dissolve cautiously 2.5 g in 25 ml of principal spot in the chromatogram obtained with the test 1 M hydrochloric acid. 12 ml of the solution complies with solution is similar in position and size to the principal limit test A for heavy metals (10 ppm). Prepare the standard spot in the chromatogram obtained with the reference using lead standard solution (1 ppm Pb) R. solution. 988 See the information section on general monographs (cover pages)