Chorionic Gonadotropin
»Chorionic Gonadotropin is a gonad-stimulating polypeptide hormone obtained from the urine of pregnant women.Its potency is not less than 1500USP Chorionic Gonadotropin Units in each mg,and not less than 80.0percent and not more than 125.0percent of the potency stated on the label.
Packaging and storage— Preserve in tight containers,preferably of Type Iglass,in a refrigerator.
USP Reference standards á11ñ— USP Endotoxin RS.USP Human Chorionic Gonadotropin RS.
Bacterial endotoxins á85ñ— It contains not more than 0.03USP Endotoxin Unit per USP Chorionic Gonadotropin Unit.
Sterility á71ñ(Where it is labeled as sterile): meets the requirements.
Acute toxicity— Select five healthy mice,weighing between 18g and 22g.Prepare a test solution as directed in the test for Pyrogen,but containing 2000USP Chorionic Gonadotropin Units per mL.Inject intravenously a dose of 0.5mLof the test solution into each of the mice.Observe the animals over the 48hours following the injection.If,at the end of 48hours,all of the animals survive and not more than one of the animals shows outward symptoms of a toxic reaction,the requirements of the test are met.If more than one of the animals show outward signs of a toxic reaction or if not more than two of the animals die,repeat the test on ten additional,similar animals:if all of the animals of the repeat test survive for 48hours and show no symptoms of a toxic reaction,the requirements of the test are met.
Water,Method Iá921ñ: not more than 5.0%.
Estrogenic activity— Dissolve a suitable quantity in saline TSto obtain a test solution containing the equivalent of 1000USP Chorionic Gonadotropin Units per mL.Into each of five rats that have been ovariectomized not less than 2weeks previously,inject subcutaneously 0.25mLof the test solution in the forenoon and in the afternoon of two successive days.On each of the three following days,take a vaginal smear from each animal:the requirements of the test are met if the cellular elements in the smears consist mainly of leucocytes,and a few nucleated epithelial cells,but no cornified epithelial cells.
Assay—
Standard preparations— Dissolve a suitable quantity of USP Human Chorionic Gonadotropin RSin a diluent consisting of saline TS,freshly prepared to contain 1mg per mLof bovine serum albumin and adjusted with sodium hydroxide TSto a pHbetween 6.9and 8.0,to obtain a solution having a known concentration of 10USP Chorionic Gonadotropin Units in each mL.Using the same diluent,prepare three Standard preparationssuch that the respective concentrations of chorionic gonadotropin constitute a geometric series such as 1:1.2:1.44or 1:2:4and such that the activity in each mLlies within the range of 0.1to 1.0Unit.
Assay preparations— Following the procedure outlined for the Standard preparations,prepare solutions of Chorionic Gonadotropin to obtain three Assay preparationscorresponding to those of the Standard.
The animals— Select 20-to 23-day-old female rats,but restrict the selection so that no rat is more than 30%heavier than the lightest.House the animals under uniform conditions of temperature,lighting,feeding,and watering.Mark the animals for identification,and divide them at random into groups of the same number but not fewer than 10animals.Assign one group to each of the three Standard preparationsand three Assay preparations,respectively.
Procedure— Inject each rat subcutaneously in the dorsal area with 0.20mLof the solution to which it was assigned,at approximately the same time on each of three consecutive days.On the afternoon of the fifth day,sacrifice the animals,and excise the uterus from each animal by cutting through the cervix,stripping off the surrounding tissue,and severing at the utero-tubal junction.Gently press out the uterine fluid on moistened absorbent paper,and weigh the uterus to the nearest 0.2mg,using a suitable balance.
Calculation— Tabulate the observed uterine weight for each rat,designated by the symbol y,for each dosage group of frats.Proceed as directed in the Assayunder Corticotropin Injection,beginning with “If the data from one or more rats.”Compute the log confidence interval L(see Confidence Intervals for Individual Assay á111ñ).If the confidence interval is more than 0.1938,which corresponds at P=0.95to confidence limits of 80%and 125%of the computed potency,repeat the assay until the combined data of two or more assays,redetermined as described under Combination of Independent Assays á111ñ,meet this limit.
Auxiliary Information— Staff Liaison:Ian DeVeau,Ph.D.,Senior Scientist
Expert Committee:(BNT)Biotechnology and Natural Therapeutics/Diagnostics
USP28–NF23Page 918
Pharmacopeial Forum:Volume No.29(6)Page 1896
Phone Number:1-301-816-8178