Diacerein

STORAGE
Protected from light.
IMPURITIES
Specified impurities : A, B, C, D.
Other detectable impurities (the following substances would,
if present at a sufcient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecied impurities and/or
by the general monograph Substances for pharmaceutical
use (2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use) : F.

EUROPEAN PHARMACOPOEIA 8.0

CHARACTERS
Appearance : yellow, crystalline powder.
Solubility : practically insoluble in water, soluble in
dimethylacetamide, slightly soluble in tetrahydrofuran,
practically insoluble in anhydrous ethanol.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : diacerein CRS.

TESTS
Impurities B and H. Liquid chromatography (2.2.29).
Carry out the test protected from light.
Solution A. Dissolve 10 g of sodium hydroxide R in 500 mL
of water R.
Solution B. Dissolve 14.7 g of sodium chloride R and 18.8 g of
glycine R in 500 mL of water R.
Solution C. Mix 25.3 volumes of solution A and 74.6 volumes
of solution B. If necessary, adjust to pH 9.5 using dilute sodium
hydroxide solution R or dilute sulfuric acid R.
Solution D. Dilute 5 mL of dilute sulfuric acid R to 500 mL
A. R = H : (2S,3R)-4-(dimethylamino)-1,2-diphenyl-3-methyl- with water R.
butan-2-ol (oxyphene),
Test solution. Dissolve 0.100 g of the substance to be examined
in 30 mL of solution A, mix for 10 min. Add 70 mL of
B. R = CO-CH3 : (1S,2R)-1-benzyl-3-(dimethylamino)-2solution B and adjust to pH 9.5 with dilute sodium hydroxide
methyl-1-phenylpropyl acetate (acetoxyphene),
solution R or dilute sulfuric acid R, if necessary. Extract with
C. R = CO-CH2-CH2-CH3 : (1S,2R)-1-benzyl-33 quantities, each of 25 mL, of methylene chloride R. Combine
(dimethylamino)-2-methyl-1-phenylpropyl butanoate
the methylene chloride extracts and wash with 2 quantities,
(butyroxyphene),
each of 8 mL, of solution C and then once with 10 mL of
solution D. Evaporate the organic layer to dryness at 33 C,
completing the drying procedure using compressed air.
Dissolve the residue in 2.0 mL of the mobile phase.
Reference solution (a). Dissolve 7.5 mg of diacerein
impurity B CRS in tetrahydrofuran R and dilute to 25.0 mL
with the same solvent. Sonicate for not more than 30 s. Dilute
1.0 mL of the solution to 100.0 mL with solution A. Dilute
5.0 mL of this solution to 50.0 mL with solution A. Mix 5.0 mL
of this solution with 25 mL of solution A for 10 min. Add
D. (1S,2S)-1-benzyl-3-(dimethylamino)-2-methyl-170 mL of solution B and adjust to pH 9.5 with dilute sodium
phenylpropyl propanoate (isopropoxyphene),
hydroxide solution R or dilute sulfuric acid R, if necessary.
Perform the extraction as described for the test solution. Care
should be taken that the time between dissolution of diacerein
impurity B in tetrahydrofuran and extraction does not exceed
30 min.
Reference solution (b). Dilute 1.0 mL of reference solution (a)
F. (2RS)-3-(dimethylamino)-2-methyl-1-phenylpropan-1to 5.0 mL with the mobile phase.
one.
Column :
size : l = 0.125 m, = 4.6 mm ;
01/2014:2409 stationary phase : irregular octadecylsilyl silica gel for
chromatography R (5 m) ;
temperature : 16 1 C.
DIACEREIN
Mobile phase : tetrahydrofuran R, acetonitrile R, 4 g/L solution
of citric acid R (8:27.5:64.5 V/V/V).
Diacereinum
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 254 nm.
Injection : 100 L.
Run time : 2.5 times the retention time of impurity B.
Retention time : impurity B = about 11 min.
System suitability : reference solution (b) :
signal-to-noise ratio : minimum 10 for the principal peak.
C19H12O8
Mr 368.3 Limit :
sum of impurities B and H : not more than the area of the
[13739-02-1]
peak due to impurity B in the chromatogram obtained with
DEFINITION
reference solution (a) (15 ppm).
4,5-Diacetoxy-9,10-dioxo-9,10-dihydroanthracene-2Related substances. Liquid chromatography (2.2.29). Carry
carboxylic acid.
out the test protected from light.
Content : 98.0 per cent to 102.0 per cent (dried substance).
Solvent mixture : mobile phase A, mobile phase B (50:50 V/V).

2028

See the information section on general monographs (cover pages)

Diacerein

EUROPEAN PHARMACOPOEIA 8.0

Test solution. Dissolve 0.100 g of the substance to be examined

in 50 mL of tetrahydrofuran R and dilute to 100.0 mL with the
solvent mixture.
Reference solution (a). Dilute 1.0 mL of the test solution
to 100.0 mL with tetrahydrofuran R. Dilute 1.0 mL of this
solution to 10.0 mL with the solvent mixture.
Reference solution (b). In order to prepare impurities D and E
in situ, add 10.0 mL of 0.01 M sodium hydroxide to 0.100 g of
the substance to be examined. Add 40 mL of tetrahydrofuran R
and dilute to 100.0 mL with the solvent mixture.
Reference solution (c). Dissolve the contents of a vial of
diacerein impurity mixture CRS (impurities C and F) in a
mixture of 0.5 mL of tetrahydrofuran R and 0.5 mL of the
solvent mixture.
Column :
size : l = 0.10 m, = 4.6 mm ;
stationary phase : end-capped polar-embedded octadecylsilyl
amorphous organosilica polymer R (5 m) ;
temperature : 30 C.
Mobile phase :
mobile phase A : to 353 mL of water R add 147 mL of
phosphoric acid R and mix ; dilute 2 mL of the solution to
1000 mL with water R ;
mobile phase B : acetonitrile R ;
Time
(min)
0-3

Mobile phase A
(per cent V/V)
80

Mobile phase B
(per cent V/V)
20

3 - 13

80 60

20 40

13 - 20

60

40

Flow rate : 1.2 mL/min.

Detection : spectrophotometer at 254 nm.
Injection : 20 L.
Identification of impurities : use the chromatogram supplied
with diacerein impurity mixture CRS and the chromatogram
obtained with reference solution (c) to identify the peaks due
to impurities C and F ; use the chromatogram obtained with
reference solution (b) to identify the peaks due to impurities D
and E.
Relative retention with reference to diacerein (retention
time = about 13.5 min) : impurity D = about 1.1 ;
impurity E = about 1.15 ; impurity C = about 1.2 ;
impurity F = about 1.3.
System suitability :
resolution : minimum 1.5 between the peaks due to
impurities D and E in the chromatogram obtained with
reference solution (b) ;
signal-to-noise ratio : minimum 100 for the principal peak
in the chromatogram obtained with reference solution (a).
Limits :
correction factors : for the calculation of content, multiply
the peak areas of the following impurities by the
corresponding correction factor : impurity C = 1.4 ;
impurity D = 1.3 ; impurity E = 1.3 ; impurity F = 9.5 ;
impurities D, E : for each impurity, not more than 5 times
the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.5 per cent) ;
impurity C : not more than twice the area of the principal
peak in the chromatogram obtained with reference
solution (a) (0.2 per cent) ;
impurity F : not more than 1.5 times the area of the principal
peak in the chromatogram obtained with reference
solution (a) (0.15 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) ;
General Notices (1) apply to all monographs and other texts

total : not more than 20 times the area of the principal peak
in the chromatogram obtained with reference solution (a)
(2.0 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).
Chromium : maximum 10 ppm.
Atomic absorption spectrometry (2.2.23, Method I).
Test solution. In a digestion bomb, dissolve 0.25 g of the
substance to be examined in a mixture of 2 mL of strong
hydrogen peroxide solution R and 6 mL of nitric acid R.
Mineralise using a microwave oven with a power-incrementing
system. Transfer quantitatively to a volumetric ask with
water R and dilute to 50.0 mL with water R. Centrifuge. Dilute
5.0 mL of the clear supernatant to 50.0 mL with water R.
Blank solution. Prepare as described for the test solution,
omitting the substance to be examined.
Stock solution. Dilute 5.0 mL of chromium standard solution
(100 ppm Cr) R to 50.0 mL with water R. Dilute 5.0 mL of
this solution to 100.0 mL with water R. Dilute 2.0 mL of this
solution to 100.0 mL with a 0.12 per cent V/V solution of
dilute nitric acid R.
Reference solutions. Prepare the reference solutions using the
stock solution, diluting with the blank solution.
Source : chromium hollow-cathode lamp using a transmission
band preferably of 0.2 nm.
Wavelength : 357.9 nm.
Atomisation device : graphite furnace.
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
1.0 g.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
related substances with the following modications.
Test solution. Dissolve 60.0 mg of the substance to be
examined in tetrahydrofuran R and dilute to 50.0 mL with the
same solvent. Dilute 2.0 mL of the solution to 25.0 mL with
the solvent mixture.
Reference solution. Dissolve 60.0 mg of diacerein CRS in
tetrahydrofuran R and dilute to 50.0 mL with the same solvent.
Dilute 2.0 mL of the solution to 25.0 mL with the solvent
mixture.
Calculate the percentage content of C19H12O8 taking into
account the assigned content of diacerein CRS.
STORAGE
In an airtight container, protected from light.
IMPURITIES
Specified impurities : B, C, D, E, F, H.
Other detectable impurities (the following substances would,
if present at a sufcient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecied impurities and/or
by the general monograph Substances for pharmaceutical
use (2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.10.
Control of impurities in substances for pharmaceutical use) : G.

B. 1,8-dihydroxy-3-(hydroxymethyl)-anthracene-9,10-dione
(aloe-emodin),

2029

Diazepam

EUROPEAN PHARMACOPOEIA 8.0

01/2008:0022

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

D. 5-acetoxy-4-hydroxy-9,10-dioxo-9,10-dihydroanthracene2-carboxylic acid (monoacetyl rhein isomer A),

E. 4-acetoxy-5-hydroxy-9,10-dioxo-9,10-dihydroanthracene2-carboxylic acid (monoacetyl rhein isomer B),

F. (10S)-3-(acetoxymethyl)-10-(2,3,4,6-tetra-O-acetyl--Dglucopyranosyl)-9-oxo-9,10-dihydroanthracene-1,8-diyl
diacetate (heptaacetyl aloin, heptaacetyl barbaloin),

G. 3-(acetoxymethyl)-10-(2,3,4,6-tetra-O-acetyl--Dglucopyranosyl)anthracene-1,8,9-triyl triacetate,

H. 3-(acetoxymethyl)-9,10-dioxo-9,10-dihydroanthracene1,8-diyl diacetate (triacetyl aloe-emodin).

2030

C16H13ClN2O
[439-14-5]

Mr 284.7

DEFINITION
7-Chloro-1-methyl-5-phenyl-1,3-dihydro-2H-1,4benzodiazepin-2-one.
Content : 99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
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.
Reference solution (a). Dilute 1.0 mL of the test solution to
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
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, = 4.6 mm ;
stationary phase : spherical end-capped octylsilyl silica gel for
chromatography R (5 m) ;
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.
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 254 nm.
Injection : 20 L.
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 (retention
time = about 9 min) : impurity E = about 0.7 ; impurity A = about
0.8 ; impurity B = about 1.3.
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.
See the information section on general monographs (cover pages)