EUROPEAN PHARMACOPOEIA 11.

0 Clobazam

Reference solution (b). Dissolve 5 mg of chlordiazepoxide CRS

and 5 mg of clonazepam CRS in the mobile phase and dilute
to 50 mL with the mobile phase. Dilute 1 mL of the solution
to 100 mL with the mobile phase.
Reference solution (c). Dilute 1.0 mL of the test solution to
200.0 mL with the mobile phase.
A. 5-chloroquinolin-8-ol, Column :
– size : l = 0.25 m, Ø = 4.6 mm,
– stationary phase : octadecylsilyl silica gel for
chromatography R (5 μm).
Mobile phase : acetonitrile R, water R (40:60 V/V).
Flow rate : 1 mL/min.
B. 5,7-dichloroquinolin-8-ol, Detection : spectrophotometer at 230 nm.
Injection : 20 μL.
Run time : 5 times the retention time of clobazam.
Retention time : clobazam = about 15 min.
System suitability : reference solution (b) :
– resolution : minimum 1.3 between the peaks due to
chlordiazepoxide and clonazepam.
C. 5,7-diiodoquinolin-8-ol. Limits :
– impurity A : not more than the area of the principal peak
in the chromatogram obtained with reference solution (a)
01/2008:1974 (0.5 per cent),
corrected 11.0 – any other impurity : not more than 0.4 times the area of
the principal peak in the chromatogram obtained with
reference solution (c) (0.2 per cent),
– total of other impurities : not more than twice the area of
the principal peak in the chromatogram obtained with
CLOBAZAM reference solution (c) (1.0 per cent),
– disregard limit : 0.1 times the area of the principal peak in
Clobazamum the chromatogram obtained with reference solution (c)
(0.05 per cent).
Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in an oven at 105 °C.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
the residue obtained in the test for loss on drying.
ASSAY
Dissolve 50.0 mg in alcohol R and dilute to 100.0 mL with the
same solvent. Dilute 2.0 mL of the solution to 250.0 mL with
C16H13ClN2O2 Mr 300.7 alcohol R. Measure the absorbance (2.2.25) at the maximum
[22316-47-8] at 232 nm.
Calculate the content of C16H13ClN2O2 taking the specific
DEFINITION
absorbance to be 1380.
7-Chloro-1-methyl-5-phenyl-1,5-dihydro-3H-1,5-
benzodiazepine-2,4-dione. IMPURITIES
Content : 97.0 per cent to 103.0 per cent (dried substance).
CHARACTERS
Appearance : white or almost white, crystalline powder.
Solubility : slightly soluble in water, freely soluble in methylene
chloride, sparingly soluble in ethanol (96 per cent).
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24). A. 7-chloro-5-phenyl-1,5-dihydro-3H-1,5-benzodiazepine-
Comparison : Ph. Eur. reference spectrum of clobazam. 2,4-dione,

TESTS
Related substances. Liquid chromatography (2.2.29).
Test solution. Dissolve 10.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 mL with the
mobile phase.
Reference solution (a). Dissolve 5.0 mg of clobazam
impurity A CRS in the mobile phase and dilute to 50.0 mL with
the mobile phase. Dilute 1.0 mL of the solution to 100.0 mL B. 1-methyl-5-phenyl-1,5-dihydro-3H-1,5-benzodiazepine-
with the mobile phase. 2,4-dione,

General Notices (1) apply to all monographs and other texts 2373
Clobetasol propionate EUROPEAN PHARMACOPOEIA 11.0

IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : clobetasol propionate CRS.
TESTS
Specific optical rotation (2.2.7) : + 112 to + 118 (dried
substance).
Dissolve 0.250 g in acetone R and dilute to 25.0 mL with the
C. (3RS)-7-chloro-1,3-dimethyl-5-phenyl-1,5-dihydro-3H- same solvent.
1,5-benzodiazepine-2,4-dione,
Related substances. Liquid chromatography (2.2.29).
Test solution (a). Dissolve 20.0 mg of the substance to be
examined in the mobile phase and dilute to 20.0 mL with the
mobile phase.
Test solution (b). Dissolve 20.0 mg of the substance to be
examined in the mobile phase and dilute to 100.0 mL with
the mobile phase.
Reference solution (a). Dissolve 20.0 mg of clobetasol
propionate CRS in the mobile phase and dilute to 100.0 mL
D. 7-chloro-1,3,3-trimethyl-5-phenyl-1,5-dihydro-3H-1,5- with the mobile phase.
benzodiazepine-2,4-dione,
Reference solution (b). Dissolve the contents of a vial of
clobetasol impurity J CRS in 2 mL of the mobile phase. To
0.5 mL of the solution add 0.5 mL of test solution (b) and
dilute to 20 mL with the mobile phase.
Reference solution (c). Dissolve the contents of a vial of
clobetasol propionate for peak identification CRS (containing
impurities A, B, D and E) in 2 mL of the mobile phase.
Reference solution (d). Dilute 1.0 mL of test solution (a) to
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
E. N-[4-chloro-2-(phenylamino)phenyl]-N-methylacetamide, to 10.0 mL with the mobile phase.
Column :
– size : l = 0.15 m, Ø = 4.6 mm ;
– stationary phase : end-capped octadecylsilyl silica gel for
chromatography R (5 μm) ;
– temperature : 30 °C.
Mobile phase : mix 10 volumes of methanol R1, 42.5 volumes
of a 7.85 g/L solution of sodium dihydrogen phosphate
monohydrate R adjusted to pH 5.5 with a 100 g/L solution
F. methyl 3-[[4-chloro-2-(phenylamino)phenyl]methyl- of sodium hydroxide R and 47.5 volumes of acetonitrile for
amino]-3-oxopropanoate. chromatography R.
Flow rate : 1.0 mL/min.
04/2020:2127 Detection : spectrophotometer at 240 nm.
Injection : 10 μL of test solution (a) and reference solutions (b),
(c) and (d).
Run time : 3 times the retention time of clobetasol propionate.
Identification of impurities: use the chromatogram obtained
CLOBETASOL PROPIONATE with reference solution (b) to identify the peak due to
impurity J ; use the chromatogram supplied with clobetasol
Clobetasoli propionas propionate for peak identification CRS and the chromatogram
obtained with reference solution (c) to identify the peaks due
to impurities A, B, D and E.
Relative retention with reference to clobetasol propionate
(retention time = about 11 min): impurity A = about 0.4 ;
impurity B = about 0.6 ; impurity J = about 1.1 ;
impurity D = about 1.2 ; impurity E = about 2.1.
System suitability : reference solution (b) :
– resolution : minimum 2.0 between the peaks due to
C25H32ClFO5 Mr 467.0 clobetasol propionate and impurity J.
[25122-46-7] Calculation of percentage contents :
DEFINITION – correction factor : multiply the peak area of impurity B by
0.6 ;
21-Chloro-9-fluoro-11β-hydroxy-16β-methyl-3,20-
dioxopregna-1,4-dien-17-yl propanoate. – for each impurity, use the concentration of clobetasol
propionate in reference solution (d).
Content : 97.0 per cent to 102.0 per cent (dried substance).
Limits :
CHARACTERS – impurities B, E: for each impurity, maximum 0.3 per cent;
Appearance : white or almost white, crystalline powder. – impurities A, D : for each impurity, maximum 0.2 per cent ;
Solubility : practically insoluble in water, freely soluble in – unspecified impurities : for each impurity, maximum
acetone, sparingly soluble in ethanol (96 per cent). 0.10 per cent ;

2374 See the information section on general monographs (cover pages)