Extraction of Ovarian Hormone From Urine.

Abstract

Since the discovery of the presence of the ovarian hormone in the blood (Loewe, 1 Frank 2 ) and in urine (Loewe 3 ), Ascheim and Zondek 4 have shown that the urines of pregnant women contain large quantities of the hormone. This finding, which has been confirmed by Laqueur, 5 Slotta 6 and others, is of exceptional value in that it provides an excellent source of the hormone from the standpoint of expense as well as availability.

Recently Zondek 7 has published a preliminary note in which he states that the common organic solvents may be used to extract the hormone. We have used several organic solvents in addition to those commonly used but have found none so satisfactory as ethyl ether. In Table I are the data obtained by 4 successive extractions of the urine with ¼ its volume of ethyl ether. The combined ethereal extracts were washed with dilute alkali and dilute acid and then distilled to dryness. The residue was taken up in hot alcohol, cooled, filtered, and the potency determined. The data of Table I indicate that this very simple procedure gives a product of considerable activity.

Owing to the danger of oxidation of the hormone by the peroxides of ethyl ether, we have made use of an alternative method which has the additional advantage of being less time-consuming. The principle involved is the familiar continuous liquid extraction of a solution but the procedure is so modified that the liquid being extracted siphons into the apparatus at the top and the spent liquid is removed at the lower end by the pressure of the column of urine and chloroform. By properly regulating the speed of ingress of the urine and the rate of boiling of the chloroform, several gallons may be run through the apparatus with practically no attention but with almost complete recovery of the hormone from the urine. The chloroform extract is washed with dilute alkali and acid and distilled. The residue which is taken up in alcohol furnishes the stock from which aqueous solutions are prepared which are considerably more refined than the alcoholic solutions.