Abstract
An improved method for the preparation of highly purified progestin is described and chemical assay methods are discussed. The extraction procedure utilizes boiling alcohol instead of hot or cold benzene. The preferred assay method entails injection of the recently castrated adult female rabbit for 5 days followed by autopsy and histological examination of the uterus though estrin-primed immature rabbits exhibit the same sensitivity to progestin as adult animals.
Highlights
THE PREPARATION OF PURIFIED PROGESTINMethods of extraction differing from the boiling neutral alcohol method previously published have been tried but only one need be given-and that only because it may eventually offer a better method of preparation than the one to be given subsequently in this paper
Since our last publication (Allen, 1930,~) giving a method for the preparation of progestin, sufficient added information has been accumulated to warrant the publication of the procedure we follow
It has since been found that three extractions are sufficient to extract all of the progestin, provided that the tissue in the bags is broken up by kneading between each extraction and the alcohol is kept boiling vigorously, and that the alcohol used for preserving the tissue by Extracting Raw Material with Benzene
Summary
Methods of extraction differing from the boiling neutral alcohol method previously published have been tried but only one need be given-and that only because it may eventually offer a better method of preparation than the one to be given subsequently in this paper. In this method, benzene was used as the extracting agent becauseof its successin the hands of Laqueur (1927) in the preparation of menformone and because of the fact that it separates the proteins from the fats readily and does away with the large amount of breakdown products formed by boiling alcohol. The one objection to benzene is, that it penetrates tissue poorly because of its relatively slight solubility in water To counteract this the fresh tissue was finely minced and ground with sea sand. The use of benzene was discontinued largely because of the irregularity of results even though it might probably with more adequate study be found to be as good as or even better than the boiling alcohol method
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