Vigabatrin
01/2020:2305 chromatogram obtained with the charged aerosol detector
VIGABATRIN
Vigabatrinum
C6H11NO2 Mr 129.2
[60643-86-9]
DEFINITION
(4RS)-4-Aminohex-5-enoic acid.
Content : 98.5 per cent to 101.5 per cent (anhydrous substance).
CHARACTERS
Appearance : white or almost white powder.
Solubility : freely soluble in water, slightly soluble in methanol, – for impurities A and B, use the concentration of each
practically insoluble in methylene chloride.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison : vigabatrin CRS.
TESTS
Related substances. Liquid chromatography (2.2.29).
Test solution. Dissolve 50.0 mg of the substance to be
examined in water R, using sonication if necessary, and dilute
to 5.0 mL with the same solvent.
Reference solution (a). Dilute 1.0 mL of the test solution to
100.0 mL with water R.
Reference solution (b). Dissolve 7.5 mg of vigabatrin
impurity A CRS and 7.5 mg of vigabatrin impurity B CRS in
water R and dilute to 50.0 mL with the same solvent. Dilute
2.0 mL of the solution to 20.0 mL with water R.
Reference solution (c). Dissolve 5.0 mg of vigabatrin
impurity E CRS and 10.0 mg of vigabatrin impurity D CRS
in water R and dilute to 50.0 mL with the same solvent. To
2.0 mL of the solution add 1.0 mL of reference solution (a)
and dilute to 20.0 mL with water R.
Reference solution (d). Dilute 2 mL of reference solution (b)
to 10 mL with water R.
Column :
– size : l = 0.10 m, Ø = 4.6 mm ;
– stationary phase : end-capped solid core phenylhexylsilyl
silica gel for chromatography R (2.7 μm) ;
– temperature : 45 °C.
Mobile phase : dissolve 2.1 g of perfluoroheptanoic acid R in a acceptance criterion for other/unspecified impurities. It
mixture of 195 mL of methanol R2 and 805 mL of water for
chromatography R.
Flow rate : 1.0 mL/min.
Post-column solution : methanol R2.
Post-column flow rate : 0.8 mL/min.
Detection : charged aerosol detector (gas source : nitrogen at
35 psi) and, for impurities A and B, spectrophotometer at
210 nm.
Autosampler : set at 15 °C.
Injection : 30 μL of the test solution and reference solutions (b),
(c) and (d).
Run time : 3 times the retention time of vigabatrin.
Identification of impurities : use the chromatogram obtained
with the spectrophotometer and with reference solution (b)
to identify the peaks due to impurities A and B ; use the
4364
www.webofpharma.com
EUROPEAN PHARMACOPOEIA 11.
and with reference solution (c) to identify the peaks due to
impurities D and E.
Relative retention with reference to vigabatrin (retention
time = about 11 min) : impurity A = about 0.3 ;
impurity E = about 0.5 ; impurity D = about 0.6 ;
impurity B = about 2.3.
System suitability :
– resolution : minimum 1.5 between the peaks due to
impurities E and D in the chromatogram obtained with
reference solution (c) ;
– signal-to-noise ratio : minimum 15 for the peak due to
vigabatrin in the chromatogram obtained with reference
solution (c); minimum 10 for the peak due to impurity B
in the chromatogram obtained with reference solution (d) ;
– symmetry factor : minimum 0.6 for the peaks due to
impurity D and vigabatrin in the chromatogram obtained
with reference solution (c) ; minimum 0.6 for the peaks due
to impurities A and B in the chromatogram obtained with
reference solution (b).
Calculation of percentage contents :
impurity in reference solution (b);
– for impurity D, use the concentration of impurity D in
reference solution (c) ;
– for impurities other than A, B and D, use the concentration
of vigabatrin in reference solution (c).
Limits :
– impurity D : maximum 0.2 per cent ;
– impurities A, B (spectrophotometer at 210 nm) : for each
impurity, maximum 0.15 per cent ;
– unspecified impurities : for each impurity, maximum
0.05 per cent ;
– total : maximum 0.5 per cent ;
– reporting threshold : 0.03 per cent.
Water (2.5.12) : maximum 0.5 per cent, determined on 0.300 g.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
1.0 g.
ASSAY
Dissolve 90 mg in 50 mL of glacial 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 12.92 mg
of C6H11NO2.
IMPURITIES
Specified impurities : A, B, D.
Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
is therefore not necessary to identify these impurities for
demonstration of compliance. See also 5.10. Control of
impurities in substances for pharmaceutical use) : E.
A. (5RS)-5-ethenylpyrrolidin-2-one,
B. (2E)-2-(2-aminoethyl)but-2-enoic acid,
D. 4-aminobutanoic acid (GABA),
See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 11.0
E. 2-[(2RS)-2-aminobut-3-en-1-yl]propanedioic acid.
01/2008:0748 at a flow rate of 40 mL/min.
corrected 11.0
VINBLASTINE SULFATE
Vinblastini sulfas
C46H60N4O13S Mr 909
[143-67-9]
DEFINITION
Vinblastine sulfate contains not less than 95.0 per
cent and not more than the equivalent of 104.0 per
cent of methyl (3aR,4R,5S,5aR,10bR,13aR)-4-
(acetyloxy)-3a-ethyl-9-[(5S,7R,9S)5-ethyl-5-hydroxy-
9-(methoxycarbonyl)-1,4,5,6,7,8,9,10-octahydro-2H-3,7-
methanoazacycloundecino[5,4-b]indol-9-yl]-5-hydroxy-
8-methoxy-6-methyl-3a,4,5,5a,6,11,12,13a-octahydro-1H-
indolizino[8,1-cd]carbazole-5-carboxylate sulfate, calculated
with reference to the dried substance.
CHARACTERS
A white or slightly yellowish, crystalline powder, very
hygroscopic, freely soluble in water, practically insoluble in
ethanol (96 per cent).
IDENTIFICATION
A. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the Ph. Eur. reference spectrum
of vinblastine sulfate.
B. Examine the chromatograms obtained in the assay. The
principal peak in the chromatogram obtained with the
test solution is similar in position and approximate size
to the principal peak in the chromatogram obtained with
reference solution (a).
TESTS
Solution S. Dissolve 50.0 mg in carbon dioxide-free water R
and dilute to 10.0 mL with the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than reference solution Y7 (2.2.2,
Method I).
pH (2.2.3). Dilute 3 mL of solution S to 10 mL with carbon
dioxide-free water R. The pH of this solution is 3.5 to 5.0.
Related substances. Examine the chromatograms obtained
in the assay. 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 (c)
(2.0 per cent) and the sum of the areas of any such peaks
is not greater than 2.5 times the area of the principal peak
in the chromatogram obtained with reference solution (c)
General Notices (1) apply to all monographs and other texts
www.webofpharma.com
Vincamine
(5.0 per cent). Disregard any peak with an area less than that
of the peak in the chromatogram obtained with reference
solution (d).
Loss on drying. Not more than 15.0 per cent, determined on
3 mg by thermogravimetry (2.2.34). Heat to 200 °C at a rate of
5 °C/min, under a stream of nitrogen for chromatography R,
ASSAY
Examine by liquid chromatography (2.2.29).
Keep the solutions in iced water before use.
Test solution. Dilute 1.0 mL of solution S (see Tests) to 5.0 mL
with water R.
Reference solution (a). Dissolve the contents of a vial of
vinblastine sulfate CRS in 5.0 mL of water R to obtain a
concentration of 1.0 mg/mL.
Reference solution (b). Dissolve 1.0 mg of vincristine
sulfate CRS in 1.0 mL of reference solution (a).
Reference solution (c). Dilute 1.0 mL of reference solution (a)
to 50.0 mL with water R.
Reference solution (d). Dilute 1.0 mL of reference solution (c)
to 20.0 mL with water 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 octylsilyl silica gel for
chromatography R (5 μm). Place between the injector and
the column a precolumn packed with suitable silica gel,
– as mobile phase at a flow rate of 1.0 mL/min a mixture of
38 volumes of a 1.5 per cent V/V solution of diethylamine R
adjusted to pH 7.5 with phosphoric acid R, 12 volumes of
acetonitrile R and 50 volumes of methanol R,
– as detector a spectrophotometer set at 262 nm,
– a loop injector.
Inject 10 μL of each solution and record the chromatograms
for 3 times the retention time of the peak due to vinblastine.
The assay is not valid unless : in the chromatogram obtained
with reference solution (b) the resolution between the peaks
due to vincristine and vinblastine is not less than 4 ; the peak
in the chromatogram obtained with reference solution (d) has
a signal-to-noise ratio not less than 5. Calculate the percentage
content of C46H60N4O13S from the area of the principal peak
in each of the chromatograms obtained with the test solution
and reference solution (a) and from the declared content of
vinblastine sulfate CRS.
STORAGE
Store in an airtight, glass container, protected from light, at a
temperature not exceeding − 20 °C. If the substance is sterile,
store in a sterile, airtight, tamper-evident glass container.
04/2021:1800
VINCAMINE
Vincaminum
C21H26N2O3 Mr 354.5
[1617-90-9]
4365