Betahistine mesilate
01/2005:1071 TESTS
BETAHISTINE MESILATE
Betahistini mesilas
C10H20N2O6S2 Mr 328.4 examined in the mobile phase and dilute to 10.0 ml with the
DEFINITION
Betahistine mesilate contains not less than 98.0 per cent
and not more than the equivalent of 101.0 per cent of
N-methyl-2-(pyridin-2-yl)ethanamine bis(methanesulphonate), phase and dilute to 50.0 ml with the mobile phase. Dilute
calculated with reference to the anhydrous, 2-propanol-free
substance.
PRODUCTION
The production method must be evaluated to determine
the potential for formation of alkyl mesilates, which is
particularly likely to occur if the reaction medium contains
lower alcohols. Where necessary, the production method
is validated to demonstrate that alkyl mesilates are not
detectable in the final product.
CHARACTERS
A white, crystalline powder, very hygroscopic, very soluble
in water, freely soluble in alcohol, very slightly soluble in
2-propanol.
IDENTIFICATION
First identification : B.
Second identification : A, C, D.
A. Melting point (2.2.14) : 108 °C to 112 °C.
B. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
betahistine mesilate CRS. Examine the substances
prepared as discs.
C. Examine by thin-layer chromatography (2.2.27), using a
suitable silica gel with a fluorescent indicator having an
optimal intensity at 254 nm as the coating substance.
Test solution. Dissolve 10 mg of the substance to be
examined in alcohol R and dilute to 2 ml with the same
solvent.
Reference solution. Dissolve 10 mg of betahistine
mesilate CRS, in alcohol R and dilute to 2 ml with the
same solvent.
Apply to the plate 2 µl of each solution. Develop over
a path of 15 cm using a mixture of 0.75 volumes of
concentrated ammonia R, 15 volumes of ethyl acetate R chromatogram obtained with reference solution (b) (0.5 per
and 30 volumes of methanol R. Dry the plate at 110 °C
for 10 min and 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 the
reference solution.
D. To 0.1 g add 5 ml of dilute hydrochloric acid R and
shake for about 5 min. Add 1 ml of barium chloride
solution R1. The solution remains clear. To a further
0.1 g add 0.5 g of anhydrous sodium carbonate R, mix
and ignite until a white residue is obtained. Allow to cool Sulphates (2.4.13). Dilute 6 ml of solution S to 15 ml with
and dissolve the residue in 7 ml of water R. The solution distilled water R. The solution complies with the limit test
gives reaction (a) of sulphates (2.3.1).
1086 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.
Solution S. Dissolve 5.0 g in carbon dioxide-free water R
prepared from distilled water R, and dilute to 50 ml with
the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and
colourless (2.2.2, Method II).
pH (2.2.3). The pH of solution S is 2.0 to 3.0.
Related substances. Examine by liquid chromatography
(2.2.29).
Test solution. Dissolve 50 mg of the substance to be
mobile phase.
Reference solution (a). Dissolve 10 mg of betahistine
mesilate CRS and 10 mg of 2-vinylpyridine R in the mobile
2.0 ml of the solution to 50.0 ml with the mobile phase.
Reference solution (b). Dilute 1.0 ml of the test solution to
100.0 ml with the mobile phase.
Reference solution (c). Dilute 2.0 ml of reference solution (b)
to 10.0 ml with the mobile phase.
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 octadecylsilyl silica gel for
chromatography R (5 µm),
— as mobile phase at a flow rate of 1 ml/min a mixture
prepared as follows : dissolve 2.0 g of sodium dodecyl
sulphate R in a mixture of 15 volumes of a 10 per cent V/V
solution of sulphuric acid R, 35 volumes of a 17 g/l
solution of tetrabutylammoniumhydrogen sulphate R
and 650 volumes of water R ; adjust to pH 3.3 using dilute
sodium hydroxide solution R and mix with 300 volumes
of acetonitrile R,
— as detector a spectrophotometer set at 260 nm.
Inject 20 µl of reference solution (a). When using a recorder,
adjust the sensitivity of the system so that the height of the
first peak in the chromatogram obtained with reference
solution (a) is not less than 70 per cent of the full scale of the
recorder. The test is not valid unless : in the chromatogram
obtained with reference solution (a), the resolution between
the peaks corresponding to 2-vinylpyridine and betahistine
mesilate is at least 3.5.
Inject 20 µl of the test solution and of reference solutions (b)
and (c). Continue the chromatography for 3 times the
retention time of betahistine mesilate (which is about 8 min).
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) (0.2 per cent) ; the sum
of the areas of any peaks, apart from the principal peak, is
not greater than half of the area of the principal peak in the
cent).
Disregard any peak with an area less than 0.025 times that
of the principal peak in the chromatogram obtained with
reference solution (b).
2-Propanol. Not more than 0.5 per cent, determined by the
test for residual solvents (2.4.24).
Chlorides (2.4.4). To 14 ml of solution S add 1 ml of water R.
The solution complies with the limit test for chlorides
(35 ppm).
for sulphates (250 ppm).
EUROPEAN PHARMACOPOEIA 5.0
Heavy metals (2.4.8). 12 ml of solution S complies with limit
test A for heavy metals (20 ppm). Prepare the standard using
lead standard solution (2 ppm Pb) R.
Water (2.5.12). Not more than 2.0 per cent, determined on
0.50 g by the semi-micro determination of water. C.
ASSAY
Dissolve 0.140 g in 50 ml of a mixture of 1 volume
of anhydrous acetic acid R and 7 volumes of acetic
anhydride 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 16.42 mg of
C10H20N2O6S2.
STORAGE
Store in an airtight container.
IMPURITIES
A. 2-ethenylpyridine.
01/2005:0312
BETAMETHASONE
Betamethasonum
C22H29FO5 Mr 392.5
DEFINITION
Betamethasone contains not less than 97.0 per cent and
not more than the equivalent of 103.0 per cent of 9-fluoro-
11β,17,21-trihydroxy-16β-methylpregna-1,4-diene-3,20-dione,
calculated with reference to the dried substance.
CHARACTERS
A white or almost white, crystalline powder, practically
insoluble in water, sparingly soluble in ethanol, very slightly
soluble in methylene chloride.
IDENTIFICATION
First identification : B, C.
Second identification : A, C, D, E.
A. Dissolve 10.0 mg in ethanol R and dilute to 100.0 ml
with the same solvent. Place 2.0 ml of the solution in a
stoppered tube, add 10.0 ml of phenylhydrazine-sulphuric
acid solution R, mix and heat in a water-bath at 60 °C for Related substances. Examine by liquid chromatography
20 min. Cool immediately. The absorbance (2.2.25) of the (2.2.29).
solution measured at 419 nm is not greater than 0.10.
B. Examine by infrared absorption spectrophotometry
(2.2.24), comparing with the spectrum obtained with
betamethasone CRS. If the spectra obtained in the solid Reference solution (a). Dissolve 2 mg of betamethasone CRS
state with the substance to be examined and the reference and 2 mg of methylprednisolone CRS in mobile phase A and
substance show differences, dissolve the substance to be dilute to 100.0 ml with the same mobile phase.
General Notices (1) apply to all monographs and other texts
Betamethasone
examined and the reference substance separately in the
smallest necessary quantity of methylene chloride R and
evaporate to dryness on a water-bath. Using the residues,
record the spectra again.
Examine by thin-layer chromatography (2.2.27), using
as the coating substance a suitable silica gel with a
fluorescent indicator having an optimal intensity at
254 nm.
Test solution. Dissolve 10 mg of the substance to be
examined in a mixture of 1 volume of methanol R and
9 volumes of methylene chloride R and dilute to 10 ml
with the same mixture of solvents.
Reference solution (a). Dissolve 20 mg of
betamethasone CRS in a mixture of 1 volume of
methanol R and 9 volumes of methylene chloride R and
dilute to 20 ml with the same mixture of solvents.
Reference solution (b). Dissolve 10 mg of
dexamethasone CRS in reference solution (a) and dilute
to 10 ml with the same solution.
Apply separately to the plate 5 µl of each solution.
Develop over a path of 15 cm using a mixture of 5 volumes
of butanol R saturated with water R, 10 volumes of
toluene R and 85 volumes of ether R. Allow the plate to
dry in air and 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). Spray with alcoholic solution of sulphuric
acid R. Heat at 120 °C for 10 min or until the spots
appear. Allow to cool. Examine the chromatograms in
daylight and in ultraviolet light at 365 nm. The principal
spot in the chromatogram obtained with the test solution
is similar in position, colour in daylight, fluorescence in
ultraviolet light at 365 nm 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 two spots which may
however not be completely separated.
D. Mix about 5 mg with 45 mg of heavy magnesium oxide R
and ignite in a crucible until an almost white residue is
obtained (usually less than 5 min). Allow to cool, add
1 ml of water R, 0.05 ml of phenolphthalein solution R1
and about 1 ml of dilute hydrochloric acid R to render
the solution colourless. Filter. Add 1.0 ml of the filtrate
to a freshly prepared mixture of 0.1 ml of alizarin S
solution R and 0.1 ml of zirconyl nitrate solution R.
Mix, allow to stand for 5 min and compare the colour of
the solution with that of a blank prepared in the same
manner. The test solution is yellow and the blank is red.
E. Add about 2 mg to 2 ml of sulphuric acid R and shake
to dissolve. Within 5 min, a deep reddish-brown colour
develops. Add the solution to 10 ml of water R and mix.
The colour is discharged and a clear solution remains.
TESTS
Specific optical rotation (2.2.7). Dissolve 0.125 g in
methanol R and dilute to 25.0 ml with the same solvent. The
specific optical rotation is + 118 to + 126, calculated with
reference to the dried substance.
Test solution. Dissolve 25.0 mg of the substance to be
examined in a mixture of equal volumes of acetonitrile R
and methanol R and dilute to 10.0 ml with the same solvent.
1087