Basal Metabolism and Other Observations Following Oestrin Injections.

Abstract

Preparation and Assay of Female Sex Hormone. The hormone used in this experiment was obtained from human pregnancy urine according to the following procedure:

Urine, made acid to Congo Red, was extracted with butyl alcohol in a continuous extractor for 48 hours. The alcoholic extract was taken to dryness, the residue dissolved in ether and the ether extracted 3 times with a saturated solution of sodium bicarbonate. The ether was removed by distillation and the residue dry-distilled under reduced pressure at 160°C. to remove butyl urethane. The residue was redissolved in ether, cooled to — 12°C. and filtered through cotton. The ether solution was taken to dryness and the residue leached out thoroughly with a 5% solution of sodium hydroxide. The alkaline solution was cooled to — 12°C. and filtered, acidified till very faintly alkaline to phenolphthalein and extracted 6 times with ether. This solution was assayed and made up for injection by pouring into olive oil and removing the ether by evaporation on a steam bath.

Assay. The method used was the vaginal smear test of Allen and Doisy. For the purpose of this experiment no smear was considered positive unless there was a complete replacement of leucocyte cells by squamous epithelial cells. The unit used is the one regularly employed in these laboratories on routine assays, namely, that amount of material which will bring 3 rats into complete oestrus providing the next higher dosage used does the same for 3 other animals. This preparation assayed as follows:

0.1 mg., E, E, E,; 0.08 mg, E, E, E; 0.06 mg., E, E, E; 0.04 mg., E, E, E; 0.02mg., E, EL, EL.

Repeat: 0.06 mg., E, E, E; 0.04 mg, E, E, E; 0.02 mg, E, E, E.∗