OXYTOCIN Oxytocinum
OXYTOCIN Oxytocinum
OXYTOCIN Oxytocinum
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EUROPEAN PHARMACOPOEIA 5.0<br />
Oxytocin bulk solution<br />
Express the content of each amino acid in moles. Calculate<br />
the relative proportions of the amino acids, taking one-sixth<br />
of the sum of the number of moles of aspartic acid, glutamic<br />
acid, proline, glycine, isoleucine and leucine as equal to one.<br />
The values fall within the following limits: aspartic acid 0.95<br />
to 1.05; glutamic acid 0.95 to 1.05; proline 0.95 to 1.05;<br />
glycine 0.95 to 1.05; leucine 0.90 to 1.10; isoleucine 0.90 to<br />
1.10; tyrosine 0.7 to 1.05; half-cystine 1.4 to 2.1; not more<br />
than traces of other amino acids are present.<br />
Related peptides. Examinebyliquidchromatography<br />
(2.2.29) as described under Assay.<br />
Inject 50 µl of the test solution. In the chromatogram<br />
obtainedtheareaofanypeak,apartfromtheprincipal<br />
peak, is not greater than 1.5 per cent of the total area of the<br />
peaks; the sum of the areas of all the peaks, apart from the<br />
principal peak, is not greater than 5 per cent of the total<br />
area of the peaks. Disregard any peak due to the solvent<br />
and any peak with an area less than 0.1 per cent of that of<br />
the principal peak.<br />
Acetic acid (2.5.34): 6.0 per cent to 10.0 per cent.<br />
Test solution. Dissolve15.0mgofthesubstancetobe<br />
examined in a mixture of 5 volumes of mobile phase B and<br />
95 volumes of mobile phase A and dilute to 10.0 ml with the<br />
same mixture of solvents.<br />
Water (2.5.12). Not more than 5.0 per cent, determined on<br />
at least 50 mg by the semi-micro determination of water.<br />
Bacterial endotoxins (2.6.14): less than 300 IU/mg, if<br />
intended for use in the manufacture of parenteral dosage<br />
forms without a further appropriate procedure for removal<br />
of bacterial endotoxins.<br />
ASSAY<br />
Examine by liquid chromatography (2.2.29).<br />
Test solution. Prepare a 0.25 mg/ml solution of the<br />
substance to be examined in a 15.6 g/l solution of sodium<br />
dihydrogen phosphate R.<br />
Reference solution. Dissolve the contents of a vial of<br />
oxytocin CRS in a 15.6 g/l solution of sodium dihydrogen<br />
phosphate R to obtain a concentration of 0.25 mg/ml.<br />
Resolution solution. Dissolve the contents of a vial of<br />
oxytocin/desmopressin validation mixture CRS in 500 µl<br />
of a 15.6 g/l solution of sodium dihydrogen phosphate R.<br />
The chromatographic procedure may be carried out using:<br />
— a stainless steel column 0.125 m long and 4.6 mm in<br />
internal diameter packed with octadecylsilyl silica gel for<br />
chromatography R (5 µm),<br />
— asmobilephaseataflowrateof1ml/min:<br />
Mobile phase A. A 15.6 g/l solution of sodium<br />
dihydrogen phosphate R,<br />
Mobile phase B. Mix 1 volume of acetonitrile for<br />
chromatography R with 1 volume of water R,<br />
Time<br />
(min)<br />
Mobile phase A<br />
(per cent V/V)<br />
Mobile phase B<br />
(per cent V/V)<br />
Comment<br />
0 - 30 70 → 40 30 → 60 linear gradient<br />
30 - 30.1 40 → 70 60 → 30 switch to initial eluent<br />
composition<br />
30.1 - 45 70 30 re-equilibration<br />
— as detector a spectrophotometer set at 220 nm.<br />
Equilibrate the column with amixtureof30volumesof<br />
mobile phase B and 70 volumes of mobile phase A.<br />
Inject 25 µl of the resolution solution. When the<br />
chromatograms are recorded in the prescribed conditions,<br />
the retention times are: oxytocin about 7.5 min and<br />
desmopressin about 10 min. The test is not valid unless the<br />
resolution between the peaks corresponding to desmopressin<br />
and oxytocin is at least 5.0.<br />
Inject 25 µl of the test solution and 25 µl of the reference<br />
solution.<br />
Calculate the content of oxytocin (C 43<br />
H 66<br />
N 12<br />
O 12<br />
S 2<br />
)fromthe<br />
peak areas in the chromatograms obtained with the test<br />
solutionandthereferencesolutionandthedeclaredcontent<br />
of C 43<br />
H 66<br />
N 12<br />
O 12<br />
S 2<br />
in oxytocin CRS.<br />
STORAGE<br />
Store in an airtight container, protected from light, at a<br />
temperature of 2 °C to 8 °C. If the substance is sterile, store<br />
in a sterile, airtight, tamper-proof container.<br />
LABELLING<br />
The label states:<br />
— the oxytocin peptide content (C 43<br />
H 66<br />
N 12<br />
O 12<br />
S 2<br />
),<br />
— where applicable, that the substance is free from bacterial<br />
endotoxins.<br />
01/2005:0779<br />
<strong>OXYTOCIN</strong> BULK SOLUTION<br />
Oxytocini solutio<br />
DEFINITION<br />
Oxytocin bulk solution is a solution of oxytocin, a cyclic<br />
nonapeptide having the structure of the hormone produced<br />
by the posterior lobe of the pituitary gland that stimulates<br />
contraction of the uterus and milk ejection in receptive<br />
mammals. It is obtained by chemical synthesis. It is available<br />
as a bulk solution with a stated concentration of not less<br />
than 0.25 mg of oxytocin per millilitre, in a solvent that<br />
may contain an appropriate antimicrobial preservative. It<br />
contains not less than 95.0 per cent and not more than<br />
105.0percentoftheamountofthepeptideC 43<br />
H 66<br />
N 12<br />
O 12<br />
S 2<br />
stated per millilitre.<br />
By convention, for the purpose of labelling oxytocin<br />
preparations, 1 mg of oxytocin peptide (C 43<br />
H 66<br />
N 12<br />
O 12<br />
S 2<br />
)is<br />
equivalent to 600 IU of biological activity.<br />
CHARACTERS<br />
A clear, colourless liquid.<br />
IDENTIFICATION<br />
Examine the chromatograms obtained under Assay. The<br />
retention time of the principal peak in the chromatogram<br />
obtained with the test solution is similar to that of the<br />
principal peak in the chromatogram obtained with the<br />
reference solution.<br />
TESTS<br />
pH (2.2.3).ThepHofthepreparationtobeexaminedis3.0<br />
to 5.0.<br />
Amino acids. Examine by means of an amino-acid analyser.<br />
Standardise the apparatus with a mixture containing<br />
equimolar amounts of ammonia, glycine and the L-form of<br />
the following amino acids:<br />
lysine threonine alanine leucine<br />
histidine serine valine tyrosine<br />
arginine glutamic acid methionine phenylalanine<br />
aspartic acid proline isoleucine<br />
GeneralNotices(1)applytoallmonographsandothertexts 2175