EUROPEAN PHARMACOPOEIA 11.
0 Vincamine
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
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(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
Vincamine EUROPEAN PHARMACOPOEIA 11.0

DEFINITION – reporting threshold : 0.05 per cent.

Methyl 14-hydroxyvincane-14β-carboxylate. Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in an oven at 105 °C.
Content : 99.0 per cent to 101.0 per cent (dried substance).
Sulfated ash (2.4.14): maximum 0.2 per cent, determined on
CHARACTERS 1.0 g.
Appearance : white or almost white, crystalline powder. ASSAY
Solubility : practically insoluble in water, soluble in methylene Dissolve 0.300 g in 30 mL of a mixture of 1 volume of acetic
chloride, slightly soluble in anhydrous ethanol. anhydride R and 5 volumes of anhydrous acetic acid R.
Titrate with 0.1 M perchloric acid, determining the end-point
IDENTIFICATION potentiometrically (2.2.20).
A. Specific optical rotation (see Tests). 1 mL of 0.1 M perchloric acid is equivalent to 35.45 mg of
C21H26N2O3.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison : vincamine CRS. IMPURITIES
Specified impurities : A, C.
TESTS
Other detectable impurities (the following substances would,
Specific optical rotation (2.2.7) : + 44.3 to + 49.0 (dried if present at a sufficient level, be detected by one or other of
substance). the tests in the monograph. They are limited by the general
Dissolve 0.1 g in dimethylformamide R and dilute to 20.0 mL acceptance criterion for other/unspecified impurities and/or
with the same solvent. by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these impurities
Related substances. Liquid chromatography (2.2.29). Prepare for demonstration of compliance. See also 5.10. Control of
the solutions immediately before use. Dissolve the samples using impurities in substances for pharmaceutical use) : B, D.
sonication, while avoiding any overheating.
Test solution. Dissolve 50.0 mg of the substance to be
examined in 10 mL of tetrahydrofuran R and dilute to
100.0 mL with the mobile phase.
Reference solution (a). Dilute 1.0 mL of the test solution to
200.0 mL with the mobile phase.
Reference solution (b). Dissolve 5 mg of vincamine for system
suitability CRS (containing impurities A, B and C) in 1 mL of
tetrahydrofuran R and dilute to 10 mL with the mobile phase.
Column : A. methyl 14-hydroxy-10-methoxyvincane-14β-carboxylate
– size : l = 0.25 m, Ø = 4.6 mm ; (10-methoxyvincamine),
– stationary phase : end-capped octadecylsilyl silica gel for
chromatography with embedded polar groups R (5 μm).
Mobile phase : tetrahydrofuran R, acetonitrile R, 15.4 g/L
solution of ammonium acetate R (17:18:65 V/V/V).
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 272 nm.
Injection : 20 μL.
B. methyl 14-hydroxyvincane-14α-carboxylate
Run time : 3.5 times the retention time of vincamine. (14-epi-vincamine),
Identification of impurities : use the chromatogram
supplied with vincamine for system suitability CRS and the
chromatogram obtained with reference solution (b) to identify
the peaks due to impurities A, B and C.
Relative retention with reference to vincamine (retention
time = about 10 min) : impurity A = about 0.8 ;
impurity B = about 0.9 ; impurity C = about 1.35.
System suitability : reference solution (b) :
C. methyl 14-hydroxy-17,18-didehydrovincane-14β-
– resolution : minimum 2.0 between the peaks due to
carboxylate (17,18-didehydrovincamine),
impurity B and vincamine.
Calculation of percentage contents :
– for each impurity, use the concentration of vincamine in
reference solution (a).
Limits :
– impurities A, C : for each impurity, maximum 0.5 per cent ;
– unspecified impurities : for each impurity, maximum
0.10 per cent ; D. methyl 14,15-didehydrovincane-14-carboxylate
– total : maximum 1.0 per cent ; (apovincamine).

4366 See the information section on general monographs (cover pages)

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