N PHARMACOPOEIA 11.

0 Cysteine hydrochloride monohydrate

01/2017:0895 10 boiling and maintain the sample on the water-bath for

corrected 10.0 1h. After cooling to room temperature reconstitute the
sample to 10 mL with water R. 2 mL of the solution gives
reaction (a) of chlorides (2.3.1).
TESTS
Solution 8. Dissolve 2.5 g in distilled water R and dilute to
STEINE HYDROCHLORIDE 50 mL with the same solvent.
¥ MONOHYDRATE Appearance of solution. The solution is clear (2.2.1) and not
propal[l,2];
more inténsely coloured than reference solution BY, (2.2.2,
ini hydrochloridum monohydricum Method IT).
Dilute 10 mL of solution S to 20 mL with water R.
보 Ny
19 , 10 Specific optical rotation (2.2.7): + 5.5 to + 7.0 (dried
'아 COH substance).
M, 1756 Dissolve 2.00 g 10 hydrochloric acid R1 and dilute to 25.0 mL
with the same acid.
Ninhydrin-positive substances. Amino acid analysis
(2.2.56). For analysis, use Method 1. Prepare the solutions
no-3-sulfanylpropanoic acid hydrochloride immediately before use.
‘The concentrations of the test solution and the reference
ermentation or of protein hydrolysis. solutions may be adapted according to the sensitivity of the
5 per cent to 101.0 per cent (dried substance). equipment used. The concentrations of all solutions are
adjusted so that the system suitability requirements described
10 general chapter 2.2.46 are fulfilled, keeping the ratios of
lite or almost white, crystalline powder or concentrations between all solutions 35 described.
stals. Solution A: dilute hydrochloric acid R1 or a sample preparation
ey soluble in water, slightly soluble 10 ethanol buffer suitable for the apparatus used.
Test solution. Dissolve 30.0 mg 아 the substance 60 be
ATION examined 10 solution A and dilute to 50.0 mL with solution A.
Reference solution (a). Dilute 1.0 mL of the test solution to
100.0 mL with solution A. Dilute 2.0 mL of this solution to
cal zotation (see Tests). 10.0 mL with solution A.
Reference solution (b). Dissolve 30.0 mg of L-cystine R
ption spectrophotometry (2.2.24).
steine hydrochloride monohydrate CRS.
(impurity A) in solution A and dilute to 100.0 mL with
solution A. Dilute 1.0 mL of the solution to 250.0 mL with
omatography (2.2.27). solution A.
solve 20 mg 아 the substance to be Reference solution (c). Dissolve 30.0 mg of proline R in
water R and dilute to 10 mL with the solution A and dilute to 100.0 mL with solution A. Dilute
Add 10 mL of a 40 g/L solution of 1.0 mL of the solution to 250.0 mL with solution A.
Rin ethanol (96 per cent) R. Allow to Reference solution (d). Dilute 6.0 mL of ammonium standard
. Dilute 2 mL of the solution 10 10 mL with
solution (100 ppm NH,) R to 50.0 mL with solution A. Dilute
1.0 mL of this solution to 100.0 mL with solution A.
ution. Dissolve 20 mg of cysteine hydrochloride Reference solution (e). Dissolve 30 mg of isoleucine R and
in water R and dilute to 10 mL with 30 mg of leucine R in solution A and dilute to 50 mL with
vent, Add 10 mL of a 40 g/L solution of
de R in ethanol (96 per cent) R. Allow to solution A. Dilute 1 mL of the solution to 200 mL with
. Dilute 2 mL of the solution 10 10 mL with solution A,
Blank solution: solution A.
a gel plate R Inject suitable, equal amounts of the test, blank and reference
acial acetic acid R, water R, butanol R solutions into the amino acid analyser. Run a program suitable
for the determination of physiological amino acids.
L, System suitability: reference solution (e):
1 2/3 of the plate. ~ resolution: minimum 1.5 between the peaks due to
isoleucine and leucine.
for 30 min,
8y With ninhydrin solution R and heat at Calculation of percentage contents:
n. - for impurity A, use the concentration of impurity A in
fincipal spot in the chromatogram obtained reference solution (b);
wolition is similar in position, colour and size - for any ninhydrin-positive substance detected at
SPotin the chromatagram obtained with 570 nm, use the concentration of cysteine hydrochloride
ution, monohydrate in reference solution (a);
05 Mg in 1 mL of dilute sodium hydroxide - for any ninhydrin-positive substance detected at 440 nm,
ImL of a 30 g/L solution of sodium use the concentration of proline in reference solution (c); if
An intense violet colour develops which a peak is above the reporting threshold at both wavelengths,
Mhish-red and then orange. Add 1 mL of use the result obtained at 570 nm for quantification.
i R. The solution becomes green. Limits:
M50 mg in 5 mL of water R. Heat to about - impurity A at 570 nm: maximum 0.5 per cent;
f€-bath and carefully add, dropwise, 5 mL of - any ninhydrin-positive substance: for each impurity,
미 Peroxide solution R. Heat the water-bath maximum 0.2 per cent;

) 0210 all monographs and other texts 2451

 Cystine EURCPEAN PHARMAGH

- total: maximum 1.0 per cent;

- reporting threshold: 0.05 per cent.
The thresholds indicated under Related substances
(Table 2034.-1) in the general monograph Substances for
pharmaceutical use (2034) do not apply.
Sulfates (2.4.13): maximum 300 ppm. CYSTINE
Dilute 10 mL of solution 10 15 mL with distilled water R.
Ammonium. Amino acid analysis (2.2.56) as described in
the test for ninhydrin-positive substances with the following Cystinum
modifications.
Injection: test solution, reference solution (d) and blank 고 NH,
g solution. 10,
kot g ’s\)\ccg«
3 Limit:
ㅇ 0욕3 H NH,

£ - ammonium at 570 nm: not more than the area of the

CiH,,N,0,8,
corresponding peak in the chromatogram obtained with [56-89-3]
reference solution (d) (0.02 per cent), taking into account
the peak due to ammonium in the chromatogram obtained
with the blank solution.
DEFINITION
Tron (2.4.9): maximum 20 ppm.
12 a separating funnel, dissolve 0.50 g in 10 mL of dilute L-Cystine (3,3"disulfanediylbis[(2R)-2-aminop
hydrochloric acid R. Shake with 3 quantitics, each of 10 mL, of acid)). :
methyl isobutyl ketonie R1, shaking for 3 min each time. 10 the Product of fermentation or of protein hydrol
combined organic layers add 10 mL of water R and shake for
3 min. Use the aqueous layer. Content: 98.5 per cent to 101.0 per cent (dri
Loss on drying (2.2.32): 8.0 per cent to 12.0 per cent,
determined 00 1.000 g by drying at a pressure not exceeding CHARACTERS
0.7 kPa for 24 h. Appearance: white or almost white, crystallis
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on Solubility: practically insoluble 10 water and i
1.0& cent). It dissolves 10 dilute solutions of alkali]
ASSAY
IDENTIFICATION
10 a ground-glass stoppered flask dissolve 0.300 g of the
substance to be examined and 4 g of potassium iodide R in First identification: A, B.
20 mL of water R. Cool the solution in iced water and add Second identification: A, C, D.
3 mL of hydrochloric acid R1 and 25.0 mL of 0.05M iodine.
Stopper the flask and allow to stand in the dark for 20 min. A. Specific optical rotation (see Tests).
Titrate with 0.1 M sodium thiosulfate using 3 mL of starch B. Infrared absorption spectrophotometry(:
solution R, added towards the end 아 the titration, as indicator.
Carry out a blank titration. Comparison: cystine CRS.
1 mL of 0.05M iodine is equivalent to 15.76 mg of C. Thin-layer chromatography (2.2.27).
C,H,CINOSS.
Test solution. Dissolve 10 mg of the substa
STORAGE examined 10 1 mL of a 103 g/L solution of
acid R and dilute to 50 mL with water R.
Protected from light.
Reference solution. Dissolve 10 mg of cystine
IMPURITIES 아 a 103 g/L solution 아 hydrochloric acid Rl
Specified impurities: A. 50 mL with water R.

Other detectable impurities (the following substances would, Plate: TLC silica gel plate R.
if present at a sufficient level, be detected by one or other of Mobile phase: concentrated ammonia R, 2-prop
the tests in the monograph. They are limited by the general (30:70 V/V).
acceptance criterion for other/unspecified impurities. It
is therefore not necessary to identify these impurities for Application: 5 11
demonstration of compliance. 866 also 5.10. Control of Development: over 2/3 of the plate.
impurities in substances for pharmaceutical use): B.
Drying: in air.
잘 빠 뇨
1020 ' 105 °C for 15 min.
“
H NH, Results: the principal spot in the chromato;
with the test solution is similar in position, 6 애
A. (2R2'R)-3,3'-disulfanediylbis(2-aminopropanoic acid) 10 the principal spot in the chromatogram 00%
(cystine), the reference solution.
1 바 D. To 0.1 g carefully add 1 mL of strong hydroget
HO. solution R and 0.1 mL of ferric chloride solution
패 10 cool. Add 1 mL of dilute hydrochloric acid
of water R. Add 1 mL of barium chloride sol
B. (25)-2-amino-3-hydroxypropanoic acid (serine). Turbidity or a white precipitate develops Wit

2452 See the information section on general monographs (&