EUROPEAN PHARMACOPOEIA 10.

0 Nitrazepam

04/2010:0415 System suitability : reference solution (b) :

– peak-to-valley ratio : minimum 4.0, where Hp = height above
the baseline of the peak due to clonazepam and Hv = height
above the baseline of the lowest point of the curve
separating this peak from the peak due to nitrazepam.
NITRAZEPAM Limits :
– unspecified impurities : for each impurity, not more than the
Nitrazepamum 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)
(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
C15H11N3O3 Mr 281.3 on 1.000 g by drying in an oven at 105 °C for 4 h.
[146-22-5] Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
1.0 g.
DEFINITION
7-Nitro-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one. ASSAY
Content : 99.0 per cent to 101.0 per cent (dried substance). Dissolve 0.250 g in 25 mL of acetic anhydride R. Titrate
with 0.1 M perchloric acid, determining the end-point
CHARACTERS potentiometrically (2.2.20).
Appearance : white or yellow, crystalline powder. 1 mL of 0.1 M perchloric acid is equivalent to 28.13 mg of
Solubility : practically insoluble in water, slightly soluble in C15H11N3O3.
ethanol (96 per cent). STORAGE
IDENTIFICATION Protected from light.
Infrared absorption spectrophotometry (2.2.24). IMPURITIES
Comparison : nitrazepam CRS. Other detectable impurities (the following substances would,
TESTS if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
Related substances. Liquid chromatography (2.2.29). Carry acceptance criterion for other/unspecified impurities and/or
out the test protected from light. by the general monograph Substances for pharmaceutical use
Test solution. Dissolve 50 mg of the substance to be examined (2034). It is therefore not necessary to identify these impurities
in acetonitrile R and dilute to 20.0 mL with the same solvent. for demonstration of compliance. See also 5.10. Control of
Reference solution (a). Dilute 1.0 mL of the test solution to impurities in substances for pharmaceutical use) : A, B, C, D.
100.0 mL with acetonitrile R. Dilute 1.0 mL of this solution to
10.0 mL with acetonitrile R.
Reference solution (b). Dissolve 2 mg of clonazepam CRS in
acetonitrile R and dilute to 100.0 mL with the same solvent.
Dilute 1.0 mL of this solution to 10.0 mL with the test solution.
Column :
– size : l = 0.25 m, Ø = 4.0 mm ;
– stationary phase : octylsilyl silica gel for chromatography R A. 3-amino-6-nitro-4-phenylquinolin-2(1H)-one,
(5 μm) ;
– temperature: 40 °C.
Mobile phase :
– mobile phase A : 7.8 g/L solution of sodium dihydrogen
phosphate R adjusted to pH 3.0 with phosphoric acid R ;
– mobile phase B : acetonitrile R ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V) B. (2-amino-5-nitrophenyl)phenylmethanone,
0-3 65 35

3 - 10 65 → 50 35 → 50

10 - 20 50 50

Flow rate : 1 mL/min.

Detection : spectrophotometer at 270 nm.
Injection : 10 μL.
Relative retention with reference to nitrazepam (retention
time = about 9 min) : clonazepam = about 1.1. C. 2-bromo-N-[4-nitro-2-(phenylcarbonyl)phenyl]acetamide,

General Notices (1) apply to all monographs and other texts 3371
Nitrendipine EUROPEAN PHARMACOPOEIA 10.0

Reference solution (c). Dilute 0.5 mL of the test solution to

20.0 mL with the mobile phase.
Reference solution (d). Mix 1.0 mL of reference solution (b)
and 1.0 mL of reference solution (c), then dilute to 25.0 mL
with the mobile phase.
Reference solution (e). Dissolve 2 mg of nitrendipine for peak
identification CRS (containing impurities B and C) in 0.5 mL
of tetrahydrofuran R and dilute to 1.0 mL with the mobile
phase.
D. 2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-N-[4-nitro-2- Column :
(phenylcarbonyl)phenyl]acetamide.
– size : l = 0.125 m, Ø = 4 mm ;
– stationary phase : irregular octadecylsilyl silica gel for
07/2012:1246 chromatography R (5 μm) ;
corrected 8.0 – temperature : 40 °C.
Mobile phase : acetonitrile R, tetrahydrofuran R, water R
([Link] V/V/V).
Flow rate : 1 mL/min.
Detection : spectrophotometer at 235 nm.
NITRENDIPINE Injection : 10 μL of the test solution and reference solutions (a),
(d) and (e).
Nitrendipinum Run time : 5 times the retention time of nitrendipine.
Identification of impurities : use the chromatogram supplied
with nitrendipine for peak identification CRS and the
chromatogram obtained with reference solution (e) to identify
the peaks due to impurities B and C ; use the chromatogram
obtained with reference solution (d) to identify the peak due
to impurity A.
Relative retention with reference to nitrendipine
(retention time = about 9 min) : impurity B = about 0.7 ;
C18H20N2O6 Mr 360.4 impurity A = about 0.8 ; impurity C = about 1.4.
[39562-70-4]
System suitability : reference solution (d) :
DEFINITION – resolution : minimum 2.0 between the peaks due to
Ethyl methyl (4RS)-2,6-dimethyl-4-(3-nitrophenyl)-1,4- impurity A and nitrendipine.
dihydropyridine-3,5-dicarboxylate. Limits :
Content : 98.5 per cent to 101.5 per cent (dried substance). – impurities B, C : for each impurity, not more than 4 times
the area of the principal peak in the chromatogram
CHARACTERS obtained with reference solution (a) (0.4 per cent) ;
Appearance : yellow, crystalline powder. – impurity A : not more than the area of the corresponding
Solubility : practically insoluble in water, freely soluble in peak in the chromatogram obtained with reference
ethyl acetate, sparingly soluble in anhydrous ethanol and in solution (d) (0.15 per cent);
methanol. – unspecified impurities : for each impurity, not more than the
It shows polymorphism (5.9). area of the principal peak in the chromatogram obtained
Exposure to ultraviolet light leads to the formation of a with reference solution (a) (0.10 per cent) ;
nitrophenylpyridine derivative. – total : maximum 0.7 per cent ;
Prepare solutions immediately before use either protected from – disregard limit : 0.5 times the area of the principal peak in
light or under long-wavelength light (> 420 nm). the chromatogram obtained with reference solution (a)
(0.05 per cent).
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24). Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in an oven at 105 °C.
Comparison : nitrendipine CRS.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
If the spectra obtained in the solid state show differences,
1.0 g.
record new spectra using 20 g/L solutions in methylene
chloride R and a 0.2 mm cell. ASSAY
TESTS Dissolve 0.160 g with gentle heating if necessary in a mixture
of 25 mL of 2-methyl-2-propanol R and 25 mL of perchloric
Related substances. Liquid chromatography (2.2.29). acid solution R. Titrate with 0.1 M cerium sulfate, using 0.1 mL
Test solution. Dissolve 20 mg of the substance to be examined of ferroin R as indicator. Titrate slowly towards the end of the
in 2.5 mL of tetrahydrofuran R and dilute to 10.0 mL with the titration. Carry out a blank titration.
mobile phase. 1 mL of 0.1 M cerium sulfate is equivalent to 18.02 mg
Reference solution (a). Dilute 1.0 mL of the test solution to of C18H20N2O6.
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
to 10.0 mL with the mobile phase. STORAGE
Reference solution (b). Dissolve 15.0 mg of nitrendipine Protected from light.
impurity A CRS in 2.5 mL of tetrahydrofuran R and dilute to
10.0 mL with the mobile phase. Dilute 1.0 mL of this solution IMPURITIES
to 20.0 mL with the mobile phase. Specified impurities : A, B, C.

3372 See the information section on general monographs (cover pages)