EUROPEAN PHARMACOPOEIA 5.

0 Betacarotene

Test solution (b). Dissolve 80.0 mg of the substance to be

examined in water R and dilute to 20.0 ml with the same
solvent.
Reference solution (a). Dissolve 50.0 mg of benzylpenicillin
sodium CRS in water R and dilute to 50.0 ml with the same
solvent.
Reference solution (b). Dissolve 10 mg of benzylpenicillin
sodium CRS and 10 mg of phenylacetic acid CRS in water R C. (2S,5R,6R)-6-[[(4-hydroxyphenyl)acetyl]amino]-3,3-
and dilute to 50 ml with the same solvent. dimethyl-7-oxo-4-thia- 1-azabicyclo[3.2.0]heptane-2-
carboxylic acid,
Reference solution (c). Dilute 1.0 ml of reference solution (a)
to 20.0 ml with water R. Dilute 1.0 ml of the solution to
50.0 ml with water R.
Reference solution (d). Dilute 4.0 ml of reference solution (a)
to 100.0 ml with water R.
The chromatographic procedure may be carried out using :
a column 0.25 m long and 4.6 mm in internal
diameter packed with octadecylsilyl silica gel for
chromatography R (5 m), D. (3S,7R,7aR)-5-benzyl-2,2-dimethyl-2,3,7,7a-
tetrahydroimidazo[5,1-b]thiazole-3,7-dicarboxylic
as mobile phase at a flow rate of 1.0 ml/min : acid (penillic acid of benzylpenicillin),
Mobile phase A. Mix 10 volumes of a 68 g/l solution of
potassium dihydrogen phosphate R adjusted to pH 3.5
with a 500 g/l solution of dilute phosphoric acid R,
30 volumes of methanol R and 60 volumes of water R.
Mobile phase B. Mix 10 volumes of a 68 g/l solution of
potassium dihydrogen phosphate R adjusted to pH 3.5
with a 500 g/l solution of dilute phosphoric acid R,
40 volumes of water R and 50 volumes of methanol R.
E. (4S)-2-[carboxy[(phenylacetyl)amino]methyl]-5,5-
as detector a spectrophotometer set at 225 nm. dimethylthiazolidine-4-carboxylic acid (penicilloic acids
Equilibrate the column with a mobile phase ratio A:B of of benzylpenicillin),
70:30. Inject 20 l of reference solution (b). The test is not
valid unless the resolution between the 2 principal peaks is
at least 6.0 (if necessary, adjust the ratio A:B of the mobile
phase) and the mass distribution ratio for the second peak
(benzylpenicillin) is 4.0 to 6.0. Inject 20 l of reference
solution (c). Adjust the system to obtain a peak with a
signal-to-noise ratio of at least 3.
F. (2RS,4S)-2-[[(phenylacetyl)amino]methyl]-5,5-
STORAGE
dimethylthiazolidine-4-carboxylic acid (penilloic acids of
Store in an airtight container. If the substance is sterile, benzylpenicillin).
store in a sterile, airtight, tamper-proof container.

LABELLING 01/2005:1069
The label states, where applicable, that the substance is free
from bacterial endotoxins. BETACAROTENE
IMPURITIES
Betacarotenum

A. (2S,5R,6R)-6-amino-3,3-dimethyl-7-oxo-4-thia-
1-azabicyclo[3.2.0]heptane-2-carboxylic acid
(6-aminopenicillanic acid),

B. phenylacetic acid, C40H56 Mr 536.9

General Notices (1) apply to all monographs and other texts 1083
Betadex
DEFINITION
Betacarotene contains not less than 96.0 per cent
and not more than the equivalent of 101.0 per cent of
(all-E)-3,7,12,16-Tetramethyl-1,18-bis(2,6,6-trimethylcyclohex-
1-enyl)octadeca-1,3,5,7,9,11,13,15,17-nonaene, calculated
with reference to the dried substance.
CHARACTERS
A brown-red or brownish-red, crystalline powder, practically
insoluble in water, slightly soluble in cyclohexane, practically
insoluble in ethanol. It is sensitive to air, heat and light,
especially in solution.
Carry out all operations as rapidly as possible avoiding
exposure to actinic light ; use freshly prepared solutions.
IDENTIFICATION
Dissolve 50.0 mg in 10 ml of chloroform R and dilute
immediately to 100.0 ml with cyclohexane R. Dilute 5.0 ml
of this solution to 100.0 ml with cyclohexane R (solution A ;
use solution A also for the test for related substances).
Dilute 5.0 ml of solution A to 50.0 ml with cyclohexane R.
(Solution B ; use solution B also for the test for related
substances and for the assay). Determine the absorbance
(2.2.25) of solution B at 455 nm and at 483 nm using
cyclohexane R as the compensation liquid. The ratio of the
absorbance at 455 nm to that at 483 nm is between 1.14
and 1.18.
TESTS
Related substances. Determine the absorbance (2.2.25) of
solution B at 455 nm and that of solution A at 340 nm, used IDENTIFICATION
in Identification. The ratio of the absorbance at 455 nm to
that at 340 nm is not less than 1.5.
Heavy metals (2.4.8). 2.0 g complies with limit test D for
heavy metals (10 ppm). Prepare the standard using 2 ml of
lead standard solution (10 ppm Pb) R.
Loss on drying (2.2.32). Not more than 0.2 per cent,
determined on 1.000 g by drying in vacuo over diphosphorus
pentoxide R at 40 C for 4 h.
Sulphated ash (2.4.14). Not more than 0.2 per cent,
determined on 1.0 g, moistened with a mixture of 2 ml of
dilute sulphuric acid R and 5 ml of alcohol R.
ASSAY
Measure the absorbance (2.2.25) of solution B used
in Identification at the maximum at 455 nm, using
cyclohexane R as the compensation liquid.
Calculate the content of C40H56 taking the specific absorbance Specific optical rotation (2.2.7) : + 160 to + 164, determined
to be 2500.
STORAGE
Store in an airtight container, protected from light, at a
temperature not exceeding 25 C.
1084 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.0
01/2005:1070
BETADEX
Betadexum
[C6H10O5]7 Mr 1135
DEFINITION
Betadex (betacyclodextrin) contains not less than 98.0 per
cent and not more than the equivalent of 101.0 per cent
of cyclo--(14)-D-heptaglucopyranoside, calculated with
reference to the dried substance.
CHARACTERS
A white or almost white, amorphous or crystalline powder,
sparingly soluble in water, freely soluble in propylene glycol,
practically insoluble in ethanol and in methylene chloride.
A. It complies with the test for specific optical rotation (see
Tests).
B. Examine the chromatograms obtained in the assay.
The retention time and size of the principal peak in
the chromatogram obtained with test solution (b) are
approximately the same as those of the principal peak in
the chromatogram obtained with reference solution (c).
C. Dissolve 0.2 g in 2 ml of iodine solution R4 by warming
on a water-bath, and allow to stand at room temperature.
A yellowish-brown precipitate is formed.
TESTS
Solution S. Dissolve 1.000 g in carbon dioxide-free water R
with heating, allow to cool and dilute to 100.0 ml with the
same solvent.
Appearance of solution. Solution S is clear (2.2.1).
pH (2.2.3). To 10 ml of solution S add 0.1 ml of a saturated
solution of potassium chloride R. The pH of the solution
is 5.0 to 8.0.
on solution S and calculated with reference to the dried
substance.
Reducing sugars
Test solution. To 1 ml of solution S add 1 ml of cupri-tartaric
solution R4. Heat on a water-bath for 10 min, cool to
room temperature. Add 10 ml of ammonium molybdate
reagent R1 and allow to stand for 15 min.