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C46H65N15O12S2 1084.24

Vasopressin, 8-L-arginine- [113-79-1].

C46H65N13O12S2 1056.22

Vasopressin, 8-L-lysine- [50-57-7].
»Vasopressin is a polypeptide hormone having the properties of causing the contraction of vascular and other smooth muscles, and of antidiuresis. It is prepared by synthesis or obtained from the posterior lobe of the pituitary of healthy, domestic animals used for food by humans. Its vasopressor activity is not less than 300 USP Vasopressin Units per mg.
Packaging and storage— Preserve in tight containers, preferably of Type I glass, in a refrigerator.
Microbial limits 61 The total bacterial count does not exceed 200 cfu per g. For products of animal origin, it meets also the requirements of the tests for absence of Salmonella species and Escherichia coli.
A: The retention time of the vasopressin peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
B: Bioidentity—About 18 hours prior to the test, select a male rat weighing between 275 g and 325 g. Inject, subcutaneously, 1 mL per kg of body weight of a solution prepared by dissolving 50 mg of phenoxybenzamine hydrochloride in 0.1 mL of alcohol, acidifying with 1 drop of hydrochloric acid, and diluting with saline TS to 5 mL. On the day of the test, anesthetize the rat, using an anesthetic substance that favors the maintenance of a uniform blood pressure. Secure the animal, and cannulate the trachea for artificial respiration. Arrange to obtain a continuous record of the blood pressure from the carotid artery. Arrange for intravenous injections by means of a suitable cannula approximately 1 mm in external diameter inserted into a femoral or jugular vein. Keep the animal warm during its preparation and during the test. Determine by trial the dose of the Standard preparation which, when injected intravenously at regular intervals of 12 to 15 minutes, will produce consistent blood pressure elevations of between 20 and 70 mm of mercury. Select the 2 doses that would be in a ratio of approximately 2 to 3, and prepare 2 doses of the Test preparation that correspond to the doses of the selected Standard preparations. Inject the rat with each dose in replicate of the Standard preparation and the Test preparation in a random fashion at regular intervals of 12 to 15 minutes, and record the blood pressures. The requirements of the test are met if the increase in blood pressure between the low dose and the high dose of the Standard preparation is comparable to that of the Test preparation.
Oxytocic activity (for product labeled of animal origin)— Proceed as directed in the Assay under Oxytocin, except that a suitable dilution of the USP Oxytocin RS will contain approximately 1.2 USP Oxytocin Units per mL of Standard preparation. The oxytocic activity of the Test preparation is not more than 1.2 USP Oxytocin Units per mL.
Ordinary impurities— The sum of the responses of impurities in the chromatogram of the Assay preparation obtained in the Assay is not more than 5% of the area of the vasopressin peak.
Residual solvents 467: meets the requirements.
(Official January 1, 2007)
Mobile phase— Dissolve 6.6 g of dibasic ammonium phosphate in about 950 mL of water, and adjust with concentrated phosphoric acid to a pH of 3.0. Dilute with water to 1 L, and mix. To 870 mL of this solution add 130 mL of acetonitrile, and mix. Filter under vacuum through a 0.45-µm nylon membrane. [NOTE—The retention time of the vasopressin peak is very sensitive to small changes in acetonitrile concentration in the Mobile phase.]
Diluent— Dissolve 5.0 g of chlorobutanol in 5.0 mL of glacial acetic acid, add 5.0 g of alcohol, 1.1 g of sodium acetate, and 1000 mL of water, and mix.
Standard preparation— Dissolve the entire contents of a vial of USP Vasopressin RS in a known volume of Diluent. [NOTE—The solution may be diluted as necessary to a working concentration range for the assay.]
Assay preparation— Transfer about 10 mg of Vasopressin, accurately weighed, to a 100-mL volumetric flask. Dissolve in 0.25% glacial acetic acid, and dilute with the same solvent to volume. Mix, and pipet 5.0 mL of this solution into a 100-mL volumetric flask, and dilute with 0.25% glacial acetic acid to volume.
Chromatographic system (see Chromatography 621)—The liquid chromatograph is equipped with a variable wavelength detector set at 220 mm and a 4.6-mm × 25-cm column that contains packing L1. The flow rate is 1.0 mL per minute. The column is allowed to equilibrate for 1 hour before making the first injection. Determine the suitability of the system (see System Suitability under Chromatography 621) as follows. Inject 20 µL of the Standard preparation into the equilibrated liquid chromatograph, allow about 60 minutes for complete elution, and record the chromatogram as directed for Procedure: the retention time of the vasopressin peak is between 6 and 9 minutes, and is completely resolved from adjacent peaks; the resolution, R, between vasopressin and the nearest adjacent peak is not less than 1.5; and the relative standard deviation for replicate injections is not more than 2.0% for vasopressin.
Procedure— Separately inject equal volumes (about 20 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the potency of Vasopressin, in USP Vasopressin Units per mg, by the formula:
20C(rU / rS)(V / W),
in which V is the volume of sample solution in which the sample was dissolved; and W is the amount, in mg, of vasopressin dissolved in the sample solution; C is the concentration in USP Vasopressin Units per mL in the Standard preparation; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information— Staff Liaison : Larry N. Callahan, Ph.D., Scientist
Expert Committee : (BBPP05) Biologics and Biotechnology - Proteins and Polysaccharides
USP29–NF24 Page 2242
Pharmacopeial Forum : Volume No. 31(4) Page 1127
Phone Number : 1-301-816-8385