Diclofenac sodium EUROPEAN PHARMACOPOEIA 5.

0
chromatogram obtained with reference solution (a). The
test is not valid unless the chromatogram obtained with
reference solution (b) shows 2 clearly separated spots.
C. Dissolve about 10 mg in 10 ml of alcohol R. To 1 ml of the
solution add 0.2 ml of a mixture, prepared immediately
before use, of equal volumes of a 6 g/l solution of
potassium ferricyanide R and a 9 g/l solution of ferric
chloride R. Allow to stand protected from light for 5 min.
Add 3 ml of a 10 g/l solution of hydrochloric acid R.
Allow to stand protected from light for 15 min. A blue
colour develops and a precipitate is formed.
D. Suspend 0.5 g of the substance to be examined in 10 ml
of water R. Stir, add water R until the substance is
dissolved. Add 2 ml of hydrochloric acid R1, stir for 1 h
and filter with the aid of vacuum. Neutralise the solution
with sodium hydroxide solution R. The solution gives
reaction (b) of potassium (2.3.1).
TESTS
Appearance of solution. Dissolve 1.25 g in methanol R and
dilute to 25.0 ml with the same solvent. The solution is clear
(2.2.1). The absorbance (2.2.25) measured at 440 nm is not
greater than 0.05.
Related substances. Examine by liquid chromatography
(2.2.29).
Test solution. Dissolve 50.0 mg of the substance to be
examined in methanol R and dilute to 50.0 ml with the same
solvent.
Reference solution (a). Dilute 2.0 ml of the test solution to
100.0 ml with methanol R. Dilute 1.0 ml of the solution to
10.0 ml with methanol R.
Reference solution (b). Dissolve 1.0 mg of diclofenac
impurity A CRS in methanol R, add 1.0 ml of the test
solution and dilute to 200.0 ml with methanol R.
The chromatographic procedure may be carried out using:
a stainless steel column 0.25 m long and 4.6 mm in
internal diameter packed with end-capped octylsilyl
silica gel for chromatography R (5 m),
as mobile phase at a flow rate of 1 ml/min a mixture
of 34 volumes of a solution containing 0.5 g/l of
phosphoric acid R and 0.8 g/l of sodium dihydrogen
phosphate R adjusted to pH 2.5 with phosphoric acid R,
and 66 volumes of methanol R,
as detector a spectrophotometer set at 254 nm.
Inject 20 l of reference solution (b). When the
chromatograms are recorded in the prescribed conditions,
the retention times are: diclofenac, about 25 min and
impurity A, about 12 min. Adjust the sensitivity of the
system so that the height of the peaks in the chromatogram
obtained is at least 50 per cent of the full scale of the
recorder. The test is not valid unless in the chromatogram
obtained the resolution between the peaks corresponding to
diclofenac and impurity A is not less than 6.5.
Inject 20 l of the test solution and 20 l of reference
solution (a). Continue the chromatography for 1.5 times the
retention time of the principal peak in the chromatogram
obtained with the test solution. In the chromatogram
obtained with the test solution: the area of any peak, apart
from the principal peak, is not greater than the area of the
principal peak in the chromatogram obtained with reference
solution (a) (0.2 per cent) ; the sum of the areas of all the
peaks, apart from the principal peak, is not greater than
2.5 times that of the principal peak in the chromatogram
obtained with reference solution (a) (0.5 per cent). Disregard
any peak with an area less than 0.25 times the area of the
principal peak in the chromatogram obtained with reference
solution (a).
Heavy metals (2.4.8). 2.0 g complies with limit test C
(10 ppm). Use a quartz crucible. Prepare the standard using
2 ml of lead standard solution (10 ppm Pb) R.
Loss on drying (2.2.32). Not more than 0.5 per cent,
determined on 1.000 g by drying in an oven at 100-105 C
for 3 h.
ASSAY
Dissolve 0.250 g in 30 ml of anhydrous acetic acid 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 33.42 mg of
C
14
H
10
Cl
2
KNO
2
.
STORAGE
In an airtight container, protected from light.
IMPURITIES
A. 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one,
B. R1 = CHO, R2 = Cl : 2-[(2,6-dichlorophenyl)amino]ben-
zaldehyde,
C. R1 = CH
2
OH, R2 = Cl : [2-[(2,6-dichlorophenyl)amino]phe-
nyl]methanol,
D. R1 = CH
2
-CO
2
H, R2 = Br: 2-[2-[(2-bromo-6-
chlorophenyl)amino]phenyl]acetic acid,
E. 1,3-dihydro-2H-indol-2-one.
01/2005:1002
DICLOFENAC SODIUM
Diclofenacum natricum
C
14
H
10
Cl
2
NNaO
2
M
r
318.1
1420 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.0 Diclofenac sodium
DEFINITION
Diclofenac sodium contains not less than 99.0 per cent and
not more than the equivalent of 101.0 per cent of sodium
2-[(2,6-dichlorophenyl)amino]phenyl]acetate, calculated with
reference to the dried substance.
CHARACTERS
A white or slightly yellowish, crystalline powder, slightly
hygroscopic, sparingly soluble in water, freely soluble in
methanol, soluble in alcohol, slightly soluble in acetone.
It melts at about 280 C, with decomposition.
IDENTIFICATION
First identification: A, D.
Second identification: B, C, D.
A. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
diclofenac sodium CRS. Examine the substances
prepared as discs.
B. Examine by thin-layer chromatography (2.2.27), using a
TLC silica gel GF
254
plate R.
Test solution. Dissolve 25 mg of the substance to be
examined in methanol R and dilute to 5 ml with the same
solvent.
Reference solution (a). Dissolve 25 mg of diclofenac
sodium CRS in methanol R and dilute to 5 ml with the
same solvent.
Reference solution (b). Dissolve 10 mg of indometacin R
in reference solution (a) and dilute to 2 ml with the same
solution.
Apply to the plate 5 l of each solution. Develop over
a path of 10 cm using a mixture of 10 volumes of
concentrated ammonia R, 10 volumes of methanol R and
80 volumes of ethyl acetate R. Allow the plate to dry in
air. Examine in ultraviolet light at 254 nm. The principal
spot in the chromatogram obtained with the test solution
is similar in position and size to the principal spot in the
chromatogram obtained with reference solution (a). The
test is not valid unless the chromatogram obtained with
reference solution (b) shows 2 clearly separated spots.
C. Dissolve about 10 mg in 10 ml of alcohol R. To 1 ml of the
solution add 0.2 ml of a mixture, prepared immediately
before use, of equal volumes of a 6 g/l solution of
potassium ferricyanide R and a 9 g/l solution of ferric
chloride R. Allow to stand protected from light for 5 min.
Add 3 ml of a 10 g/l solution of hydrochloric acid R.
Allow to stand, protected from light, for 15 min. A blue
colour develops and a precipitate is formed.
D. Dissolve 60 mg in 0.5 ml of methanol R and add 0.5 ml of
water R. The solution gives reaction (b) of sodium (2.3.1).
TESTS
Appearance of solution. Dissolve 1.25 g in methanol R and
dilute to 25.0 ml with the same solvent. The solution is clear
(2.2.1). The absorbance (2.2.25) measured at 440 nm is not
greater than 0.05.
Related substances. Examine by liquid chromatography
(2.2.29).
Test solution. Dissolve 50.0 mg of the substance to be
examined in methanol R and dilute to 50.0 ml with the same
solvent.
Reference solution (a). Dilute 2.0 ml of the test solution to
100.0 ml with methanol R. Dilute 1.0 ml of the solution to
10.0 ml with methanol R.
Reference solution (b). Dissolve 1.0 mg of diclofenac
impurity A CRS in methanol R, add 1.0 ml of the test
solution and dilute to 200.0 ml with methanol R.
The chromatographic procedure may be carried out using:
a stainless steel column 0.25 m long and 4.6 mm in
internal diameter packed with end-capped octylsilyl
silica gel for chromatography R (5 m),
as mobile phase at a flow rate of 1 ml/min a mixture
of 34 volumes of a solution containing 0.5 g/l of
phosphoric acid R and 0.8 g/l of sodium dihydrogen
phosphate R adjusted to pH 2.5 with phosphoric acid R,
and 66 volumes of methanol R,
as detector a spectrophotometer set at 254 nm.
Inject 20 l of reference solution (b). When the
chromatograms are recorded in the prescribed conditions,
the retention times are about 25 min for diclofenac and about
12 min for impurity A. Adjust the sensitivity of the system so
that the height of the peaks in the chromatogram obtained
with reference solution (b) is not less than 50 per cent of
the full scale of the recorder. Continue the chromatography
for 1.5 times the retention time of diclofenac. The test
is not valid unless in the chromatogram obtained the
resolution between the peaks corresponding to diclofenac
and impurity A is at least 6.5.
Inject 20 l of the test solution and 20 l of reference
solution (a). In the chromatogram obtained with the test
solution: the area of any peak, apart from the principal
peak, is not greater than the area of the principal peak in the
chromatogram obtained with reference solution (a) (0.2 per
cent) ; the sum of the areas of all the peaks apart from the
principal peak is not greater than 2.5 times that of the
principal peak in the chromatogram obtained with reference
solution (a) (0.5 per cent). Disregard any peak with an area
less than 0.25 times the area of the principal peak in the
chromatogram obtained with reference solution (a).
Heavy metals (2.4.8). 2.0 g complies with limit test C
(10 ppm). Use a quartz crucible. Prepare the standard using
2 ml of lead standard solution (10 ppm Pb) R.
Loss on drying (2.2.32). Not more than 0.5 per cent,
determined on 1.000 g by drying in an oven at 100-105 C
for 3 h.
ASSAY
Dissolve 0.250 g in 30 ml of anhydrous acetic acid 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 31.81 mg of
C
14
H
10
Cl
2
NNaO
2
.
STORAGE
In an airtight container, protected from light.
IMPURITIES
A. 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one,
General Notices (1) apply to all monographs and other texts 1421
Dicloxacillin sodium EUROPEAN PHARMACOPOEIA 5.0
B. R1 = CHO, R2 = Cl : 2-[(2,6-dichlorophenyl)amino]ben-
zaldehyde,
C. R1 = CH
2
OH, R2 = Cl : [2-[(2,6-dichlorophenyl)amino]phe-
nyl]methanol,
D. R1 = CH
2
-CO
2
H, R2 = Br: 2-[2-[(2-bromo-6-
chlorophenyl)amino]phenyl]acetic acid,
E. 1,3-dihydro-2H-indol-2-one.
01/2005:0663
DICLOXACILLIN SODIUM
Dicloxacillinum natricum
C
19
H
16
Cl
2
N
3
NaO
5
S,H
2
O M
r
510.3
DEFINITION
Dicloxacillin sodium contains not less than 95.0 per
cent and not more than the equivalent of 101.0 per
cent of sodium (2S,5R,6R)-6-[[[3-(2,6-dichlorophenyl)-5-
methylisoxazol-4-yl]carbonyl]amino]-3,3-dimethyl-7-oxo-4-
thia-1-azabicyclo[3.2.0]heptane-2-carboxylate, calculated
with reference to the anhydrous substance.
CHARACTERS
A white or almost white, crystalline powder, hygroscopic,
freely soluble in water, soluble in alcohol and in methanol.
IDENTIFICATION
First identification: A, D.
Second identification: B, C, D.
A. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
dicloxacillin sodium CRS. Examine the substances
prepared as discs.
B. Examine by thin-layer chromatography (2.2.27), using
silanised silica gel H R as the coating substance.
Test solution. Dissolve 25 mg of the substance to be
examined in 5 ml of water R.
Reference solution (a). Dissolve 25 mg of dicloxacillin
sodium CRS in 5 ml of water R.
Reference solution (b). Dissolve 25 mg each of
cloxacillin sodium CRS, dicloxacillin sodium CRS and
flucloxacillin sodium CRS in 5 ml of water R.
Apply to the plate 1 l of each solution. Develop over a
path of 15 cm using a mixture of 30 volumes of acetone R
and 70 volumes of a 154 g/l solution of ammonium
acetate R the pH of which has been adjusted to 5.0 with
glacial acetic acid R. Allow the plate to dry in air and
expose it to iodine vapour until the spots appear. Examine
in daylight. The principal spot in the chromatogram
obtained with the test solution is similar in position,
colour and size to the principal spot in the chromatogram
obtained with reference solution (a). The test is not
valid unless the chromatogram obtained with reference
solution (b) shows three clearly separated spots.
C. Place about 2 mg in a test-tube about 150 mm long and
15 mm in diameter. Moisten with 0.05 ml of water R and
add 2 ml of sulphuric acid-formaldehyde reagent R. Mix
the contents of the tube by swirling; the colour of the
solution is slightly greenish-yellow. Place the test-tube in
a water-bath for 1 min; a yellow colour develops.
D. It gives reaction (a) of sodium (2.3.1).
TESTS
Solution S. Dissolve 2.50 g in carbon dioxide-free water R
and dilute to 25.0 ml with the same solvent.
Appearance of solution. Solution S is clear (2.2.1). The
absorbance (2.2.25) of solution S measured at 430 nm is
not greater than 0.04.
pH (2.2.3). The pH of solution S is 5.0 to 7.0.
Specific optical rotation (2.2.7). Dissolve 0.250 g in water R
and dilute to 25.0 ml with the same solvent. The specific
optical rotation is + 128 to + 143, calculated with reference
to the anhydrous substance.
Related substances. Examine by liquid chromatography
(2.2.29) as prescribed under Assay. Inject reference
solution (b). Adjust the sensitivity of the system so that the
height of the principal peak in the chromatogram obtained
is at least 50 per cent of the full scale of the recorder.
Inject test solution (a) and continue the chromatography
for five times the retention time of the principal peak. In
the chromatogram obtained with test solution (a) : the area
of any peak, apart from the principal peak, is not greater
than the area of the principal peak in the chromatogram
obtained with reference solution (b) (1 per cent) ; the sum of
the areas of all peaks, apart from the principal peak, is not
greater than five times the area of the principal peak in the
chromatogram obtained with reference solution (b) (5 per
cent). Disregard any peak with an area less than 0.05 times
the area of the principal peak in the chromatogram obtained
with reference solution (b).
N,N-Dimethylaniline (2.4.26, Method B). Not more than
20 ppm.
2-Ethylhexanoic acid (2.4.28). Not more than 0.8 per
cent m/m.
Water (2.5.12) : 3.0 per cent to 4.5 per cent, determined on
0.300 g by the semi-micro determination of water.
Pyrogens (2.6.8). If intended for use in the manufacture
of parenteral dosage forms without a further appropriate
procedure for the removal of pyrogens, it complies with the
test for pyrogens. Inject per kilogram of the rabbits mass
1 ml of a solution in water for injections R containing 20 mg
of the substance to be examined per millilitre.
ASSAY
Examine by liquid chromatography (2.2.29).
Test solution (a). Dissolve 50.0 mg of the substance to be
examined in the mobile phase and dilute to 50.0 ml with the
mobile phase.
1422 See the information section on general monographs (cover pages)