Ammonium glycyrrhizate

EUROPEAN PHARMACOPOEIA 5.0

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

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.

DEFINITION
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.

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),

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

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.

IMPURITIES

STORAGE
Store in an airtight container.
01/2005:0594

AMOBARBITAL
Amobarbitalum

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

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

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