EUROPEAN PHARMACOPOEIA 11.

0 Diazepam

01/2017:0022

C. 4,5-dihydroxy-9,10-dioxo-9,10-dihydroanthracene-2- DIAZEPAM
carboxylic acid (rhein),
Diazepamum

D. 5-(acetyloxy)-4-hydroxy-9,10-dioxo-9,10-dihydro-
anthracene-2-carboxylic acid (monoacetyl rhein isomer A),
C16H13ClN2O Mr 284.7
[439-14-5]
DEFINITION
7-Chloro-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-
benzodiazepin-2-one.
Content : 99.0 per cent to 101.0 per cent (dried substance).

E. 4-(acetyloxy)-5-hydroxy-9,10-dioxo-9,10-dihydro- CHARACTERS
anthracene-2-carboxylic acid (monoacetyl rhein isomer B), Appearance : white or almost white, crystalline powder.
Solubility : very slightly soluble in water, soluble in ethanol
(96 per cent).
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : diazepam CRS.
TESTS
Related substances. Liquid chromatography (2.2.29). Prepare
the solutions protected from bright light.
Test solution. Dissolve 25.0 mg of the substance to be
examined in 0.5 mL of acetonitrile R and dilute to 50.0 mL
with the mobile phase.
F. (10S)-3-[(acetyloxy)methyl]-10-(2,3,4,6-tetra-O-acetyl-β- Reference solution (a). Dilute 1.0 mL of the test solution to
D-glucopyranosyl)-9-oxo-9,10-dihydroanthracene-1,8-diyl 100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
diacetate (heptaacetyl aloin, heptaacetyl barbaloin), to 10.0 mL with the mobile phase.
Reference solution (b). Dissolve the contents of a vial of
diazepam for system suitability CRS (containing impurities A,
B and E) in 1.0 mL of the mobile phase.
Column :
– size : l = 0.15 m, Ø = 4.6 mm ;
– stationary phase : spherical end-capped octylsilyl silica gel for
chromatography R (5 μm) ;
– temperature : 30 °C.
Mobile phase : mix 22 volumes of acetonitrile R, 34 volumes of
methanol R and 44 volumes of a 3.4 g/L solution of potassium
dihydrogen phosphate R previously adjusted to pH 5.0 with
dilute sodium hydroxide solution R.
G. 3-[(acetyloxy)methyl]-10-(2,3,4,6-tetra-O-acetyl-β-D- Flow rate : 1.0 mL/min.
glucopyranosyl)anthracene-1,8,9-triyl triacetate, Detection : spectrophotometer at 254 nm.
Injection : 20 μL.
Run time : about 4 times the retention time of diazepam.
Identification of impurities : use the chromatogram
supplied with diazepam for system suitability CRS and the
chromatogram obtained with reference solution (b) to identify
the peaks due to impurities A, B and E.
Relative retention with reference to diazepam
H. 3-[(acetyloxy)methyl]-9,10-dioxo-9,10-dihydro- (retention time = about 9 min) : impurity E = about 0.7 ;
anthracene-1,8-diyl diacetate (triacetyl aloe-emodin). impurity A = about 0.8 ; impurity B = about 1.3.

General Notices (1) apply to all monographs and other texts 2513
Diazoxide EUROPEAN PHARMACOPOEIA 11.0

System suitability : reference solution (b) :

– resolution : minimum 2.5 between the peaks due to
impurities E and A and minimum 6.0 between the peaks
due to impurity A and diazepam.
Limits :
– correction factors : for the calculation of content,
multiply the peak areas of the following impurities by
the corresponding correction factor : impurity B = 1.3 ; C. 3-amino-6-chloro-1-methyl-4-phenylquinolin-2(1H)-one,
impurity E = 1.3 ;
– impurities A, B, E : for each impurity, not more than the
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.1 per cent) ;
– unspecified impurities : for each impurity, not more than the
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.10 per cent) ;
– total : not more than twice the area of the principal peak
in the chromatogram obtained with reference solution (a) D. [5-chloro-2-(methylamino)phenyl]phenylmethanone,
(0.2 per cent) ;
– disregard limit : 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
(0.05 per cent).
Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in vacuo at 60 °C for 4 h.
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g.
E. 6-chloro-1-methyl-4-phenylquinazolin-2(1H)-one,
ASSAY
Dissolve 0.200 g in 50 mL of acetic anhydride R. Titrate
with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.2.20).
1 mL of 0.1 M perchloric acid is equivalent to 28.47 mg
of C16H13ClN2O.
STORAGE
F. 7-chloro-2-methoxy-5-phenyl-3H-1,4-benzodiazepine.
Protected from light.
IMPURITIES 01/2008:0550
Specified impurities : A, B, E. corrected 11.0
Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use DIAZOXIDE
(2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.10. Control of Diazoxidum
impurities in substances for pharmaceutical use) : C, D, F.

C8H7ClN2O2S Mr 230.7
[364-98-7]
DEFINITION
A. 7-chloro-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2- Diazoxide contains not less than 98.0 per cent and
one (nordazepam), not more than the equivalent of 101.0 per cent of
7-chloro-3-methyl-2H-1,2,4-benzothiadiazine 1,1-dioxide,
calculated with reference to the dried substance.
CHARACTERS
A white or almost white, fine or crystalline powder, practically
insoluble in water, freely soluble in dimethylformamide,
slightly soluble in ethanol (96 per cent). It is very soluble in
dilute solutions of the alkali hydroxides.
IDENTIFICATION
B. N-(2-benzoyl-4-chlorophenyl)-2-chloro-N- First identification : B.
methylacetamide, Second identification : A, C, D.

2514 See the information section on general monographs (cover pages)