Carbidopa EUROPEAN PHARMACOPOEIA 11.

01/2017:0755 Test solution (b). Place 25 g of strongly basic anion-exchange

corrected 9.4 resin R into each of 2 conical flasks with ground-glass stoppers.
To each, add 150 mL of carbon dioxide-free water R and shake
from time to time during 30 min. Decant the liquid from both
flasks and repeat the process with further quantities, each of
150 mL, of carbon dioxide-free water R.
Take two 100 mL measuring cylinders 3.5-4.5 cm in internal
CARBIDOPA diameter and label these A and B. Into cylinder A, transfer
as completely as possible the resin from 1 conical flask using
Carbidopum 60 mL of carbon dioxide-free water R ; into cylinder B, transfer
the 2nd quantity of resin, this time using 20 mL of carbon
dioxide-free water R.
Into each cylinder, insert a gas-inlet tube, the end of which has
an internal diameter of 2-3 mm and which reaches almost to
the bottom of the cylinder. Pass a rapid stream of nitrogen for
chromatography R through each mixture so that homogeneous
C10H14N2O4,H2O Mr 244.2 suspensions are formed. After 30 min, without interrupting
[38821-49-7] the gas flow, add 1.0 mL of test solution (a) to cylinder A ;
DEFINITION after 1 min stop the gas flow into cylinder A and transfer the
(2S)-3-(3,4-Dihydroxyphenyl)-2-hydrazino-2- contents, through a moistened filter paper, into cylinder B.
methylpropanoic acid monohydrate. After 1 min, stop the gas flow to cylinder B and pour the
solution immediately through a moistened filter paper into
Content : 98.5 per cent to 101.0 per cent (dried substance). a freshly prepared mixture of 1 mL of a 200 g/L solution
CHARACTERS of salicylaldehyde R in methanol R and 20 mL of phosphate
buffer solution pH 5.5 R in a conical flask ; shake thoroughly
Appearance : white or yellowish-white powder. for 1 min and heat in a water-bath at 60 °C for 15 min. The
Solubility : slightly soluble in water, very slightly soluble in liquid becomes clear. Allow to cool, add 2.0 mL of toluene R
ethanol (96 per cent), practically insoluble in methylene and shake vigorously for 2 min. Transfer the mixture into a
chloride. It dissolves in dilute solutions of mineral acids. centrifuge tube and centrifuge.
IDENTIFICATION Separate the toluene layer in a 100 mL separating funnel and
shake vigorously with 2 quantities, each of 20 mL, of a 200 g/L
First identification : A, C. solution of sodium metabisulfite R and finally with 2 quantities,
Second identification : A, B, D, E. each of 50 mL, of water R. Separate the toluene layer.
A. Specific optical rotation (see Tests). Reference solution (a). Dissolve 10 mg of hydrazine sulfate R in
B. Ultraviolet and visible absorption spectrophotometry dilute hydrochloric acid R and dilute to 50 mL with the same
(2.2.25). acid. Dilute 1.0 mL of this solution to 10.0 mL with dilute
Test solution. Dissolve 50 mg in an 8.5 g/L solution of hydrochloric acid R.
hydrochloric acid R in methanol R and dilute to 100.0 mL Reference solution (b). Prepare the solution at the same time
with the same solution. Dilute 10.0 mL of this solution to and in the same manner as described for test solution (b)
100.0 mL with an 8.5 g/L solution of hydrochloric acid R in using 1.0 mL of reference solution (a) instead of 1.0 mL of
methanol R. test solution (a).
Spectral range : 230-350 nm. Plate : TLC silanised silica gel plate R.
Absorption maximum : at 283 nm. Mobile phase : water R, methanol R (10:20 V/V).
Specific absorbance at the absorption maximum : 135 to 150 Application : 10 μL of test solution (b) and reference
(dried substance). solution (b).
C. Infrared absorption spectrophotometry (2.2.24). Development : over 1/2 of the plate.
Comparison : carbidopa CRS. Drying : in air.
D. Shake vigorously about 5 mg with 10 mL of water R for Detection : examine in ultraviolet light at 365 nm.
1 min and add 0.3 mL of ferric chloride solution R2. An Limit :
intense green colour is produced, which quickly changes to – hydrazine : any spot showing a yellow fluorescence is
reddish-brown. not more intense than the corresponding spot in the
E. Suspend about 20 mg in 5 mL of water R and add 5 mL chromatogram obtained with reference solution (b)
of cupri-tartaric solution R. On heating, the colour of the (20 ppm).
solution changes to dark brown and a red precipitate is
formed. Related substances. Liquid chromatography (2.2.29). Prepare
the solutions immediately before use.
TESTS Buffer solution. Dissolve 6.9 g of sodium dihydrogen phosphate
Appearance of solution. The solution is clear (2.2.1) and not monohydrate R in 900 mL of water for chromatography R,
more intensely coloured than reference solution BY6 or B6 adjust to pH 2.2 with phosphoric acid R and dilute to 1000 mL
(2.2.2, Method II). with water for chromatography R.
Dissolve 0.25 g in 25 mL of 1 M hydrochloric acid. Test solution. Dissolve 20.0 mg of the substance to be
examined in the mobile phase, add 20 μL of hydrochloric
Specific optical rotation (2.2.7) : − 26.5 to − 22.5 (dried acid R and dilute to 10.0 mL with the mobile phase.
substance).
Reference solution (a). Dilute 1.0 mL of the test solution to
Using an ultrasonic bath, dissolve completely 0.250 g in 100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
aluminium chloride solution R and dilute to 25.0 mL with the to 10.0 mL with the mobile phase.
same solution.
Reference solution (b). Dissolve 4 mg of carbidopa for system
Hydrazine. Thin-layer chromatography (2.2.27). suitability CRS (containing impurities A, D, E, I and J) in the
Test solution (a). Dissolve 0.50 g in dilute hydrochloric acid R mobile phase, add 4 μL of hydrochloric acid R and dilute to
and dilute to 2.0 mL with the same acid. 2.0 mL with the mobile phase.

2202 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 11.0 Carbidopa

Reference solution (c). Dissolve the contents of a vial of acceptance criterion for other/unspecified impurities and/or
carbidopa impurity mixture CRS (impurities F and H) in by the general monograph Substances for pharmaceutical use
1.0 mL of reference solution (b). (2034). It is therefore not necessary to identify these impurities
Column : for demonstration of compliance. See also 5.10. Control of
– size : l = 0.15 m, Ø = 4.6 mm ; impurities in substances for pharmaceutical use): B, C, G.
– stationary phase : end-capped ethylene-bridged octadecylsilyl
silica gel for chromatography (hybrid material) R (3.5 μm) ;
– temperature : 25 °C.
Mobile phase : ethanol (96 per cent) R, buffer solution
(7:93 V/V).
Flow rate : 1.0 mL/min. A. (2S)-2-amino-3-(3,4-dihydroxyphenyl)-2-methylpropanoic
acid (L-methyldopa),
Detection : spectrophotometer at 280 nm.
Injection : 20 μL.
Run time : 6 times the retention time of carbidopa.
Identification of impurities : use the chromatogram obtained
with reference solution (c) to identify the peaks due to
impurities A, D + E, F, H, I and J.
Relative retention with reference to carbidopa B. methyl (2S)-2-amino-3-(3,4-dihydroxyphenyl)-2-
(retention time = about 3 min): impurity A = about methylpropanoate,
0.9 ; impurities D and E = about 1.9 ; impurity H = about 2.5 ;
impurity I = about 3.7 ; impurity J = about 4.0 ;
impurity F = about 4.4.
System suitability :
– resolution : minimum 1.5 between the peaks due to
impurity A and carbidopa in the chromatogram obtained
with reference solution (b); minimum 1.5 between the C. (2S)-2-hydrazino-3-(4-hydroxy-3-methoxyphenyl)-2-
peaks due to impurities I and J in the chromatogram methylpropanoic acid (3-O-methylcarbidopa),
obtained with reference solution (b);
– signal-to-noise ratio : minimum 30 for the principal peak in
the chromatogram obtained with reference solution (a).
Calculation of percentage contents :
– correction factors : multiply the peak areas of the
following impurities by the corresponding correction
factor : impurities D and E = 1.5 ; impurity I = 1.5 ;
impurity J = 1.5 ;
– for each impurity, use the concentration of carbidopa in D. methyl (2S)-2-(2-cyclohexylidenehydrazino)-3-(3,4-
reference solution (a). dihydroxyphenyl)-2-methylpropanoate,
Limits :
– impurity A : maximum 0.5 per cent ;
– impurity J : maximum 0.25 per cent ;
– sum of impurities D and E : maximum 0.2 per cent ;
– impurities F, H, I : for each impurity, maximum 0.15 per
cent ;
– unspecified impurities : for each impurity, maximum E. methyl (2S)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-
0.10 per cent ; methylpropanoate,
– total : maximum 1.0 per cent ;
– reporting threshold : 0.05 per cent.
Loss on drying (2.2.32): 6.9 per cent to 7.9 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.
F. ethyl (2S)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2-
ASSAY methylpropanoate,
Dissolve 0.150 g with gentle heating in 75 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 22.62 mg
of C10H14N2O4.
G. 1-(3,4-dihydroxyphenyl)propan-2-one,
STORAGE
Protected from light.
IMPURITIES
Specified impurities : A, D, E, F, H, I, J.
Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of H. (2S)-2-hydrazino-3-(3-hydroxy-4-methoxyphenyl)-2-
the tests in the monograph. They are limited by the general methylpropanoic acid,

General Notices (1) apply to all monographs and other texts 2203
Carbimazole EUROPEAN PHARMACOPOEIA 11.0

D. Dissolve about 10 mg in a mixture of 0.05 mL of dilute

hydrochloric acid R and 50 mL of water R. Add 1 mL of
potassium iodobismuthate solution R. A red precipitate is
formed.

I. (2S)-3-(3-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2- TESTS
methylpropanoic acid, Related substances. Liquid chromatography (2.2.29). Prepare
the solutions immediately before use.
Solvent mixture : acetonitrile R, water R (20:80 V/V).
Test solution. Dissolve 25.0 mg of the substance to be
examined in the solvent mixture and dilute to 50.0 mL with
the solvent mixture.
Reference solution (a). Dissolve 5 mg of thiamazole CRS
J. (2S)-3-(2-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2- (impurity A) in the solvent mixture and dilute to 100.0 mL
methylpropanoic acid. with the solvent mixture. Mix 1 mL of the solution with 2 mL
of the test solution and dilute to 10.0 mL with the solvent
mixture.
04/2014:0884 Reference solution (b). Dissolve 5.0 mg of thiamazole CRS
corrected 10.0 (impurity A) in the solvent mixture and dilute to 100.0 mL
with the solvent mixture. Dilute 1.0 mL of the solution to
50.0 mL with the solvent mixture.
Reference solution (c). Dilute 1.0 mL of the test solution to
100.0 mL with the solvent mixture. Dilute 1.0 mL of this
solution to 10.0 mL with the solvent mixture.
CARBIMAZOLE
Reference solution (d). Dissolve 25.0 mg of carbimazole CRS
in the solvent mixture and dilute to 50.0 mL with the solvent
Carbimazolum mixture.
Column :
– size : l = 0.15 m, Ø = 3.9 mm ;
– stationary phase : end-capped octadecylsilyl silica gel for
chromatography R (5 μm).
C7H10N2O2S Mr 186.2 Mobile phase : acetonitrile R, water R (10:90 V/V).
[22232-54-8]
Flow rate : 1 mL/min.
DEFINITION Detection : spectrophotometer at 254 nm.
Ethyl 3-methyl-2-thioxo-2,3-dihydro-1H-imidazole-1- Injection : 10 μL of the test solution and reference solutions (a),
carboxylate. (b) and (c).
Content : 98.0 per cent to 102.0 per cent (dried substance). Run time : 1.5 times the retention time of carbimazole.
CHARACTERS Identification of impurities : use the chromatogram obtained
with reference solution (a) to identify the peak due to
Appearance : white or yellowish-white, crystalline powder.
impurity A.
Solubility : slightly soluble in water, soluble in acetone and in
ethanol (96 per cent). Relative retention with reference to carbimazole (retention
time = about 6 min) : impurity A = about 0.2.
IDENTIFICATION System suitability : reference solution (a) :
First identification : B. – resolution : minimum 5.0 between the peaks due to
Second identification : A, C, D. impurity A and carbimazole.
A. Melting point (2.2.14) : 122 °C to 125 °C. Limits :
B. Infrared absorption spectrophotometry (2.2.24). – impurity A : not more than 2.5 times the area of the
Comparison : carbimazole CRS. principal peak in the chromatogram obtained with
reference solution (b) (0.5 per cent) ;
C. Thin-layer chromatography (2.2.27).
– unspecified impurities : for each impurity, not more than the
Test solution. Dissolve 10 mg of the substance to be area of the principal peak in the chromatogram obtained
examined in methylene chloride R and dilute to 10.0 mL with reference solution (c) (0.10 per cent) ;
with the same solvent.
– total : maximum 0.6 per cent ;
Reference solution. Dissolve 10 mg of carbimazole CRS in
methylene chloride R and dilute to 10.0 mL with the same – disregard limit : 0.5 times the area of the principal peak in
solvent. the chromatogram obtained with reference solution (c)
(0.05 per cent).
Plate : TLC silica gel F254 plate R.
Mobile phase : acetone R, methylene chloride R (20:80 V/V). Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in a desiccator at a pressure not exceeding
Application : 10 μL. 0.7 kPa for 24 h.
Development : over 3/4 of the plate. Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
Drying : in air for 30 min. 1.0 g.
Detection : examine in ultraviolet light at 254 nm.
ASSAY
Results : the principal spot in the chromatogram obtained
with the test solution is similar in position and size to the Liquid chromatography (2.2.29) as described in the test for
principal spot in the chromatogram obtained with the related substances with the following modification.
reference solution. Injection : test solution and reference solution (d).

2204 See the information section on general monographs (cover pages)