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The purpose of this report is to present evidence obtained by gas chromatography combined with mass spectroscopy that pregn-4-ene-3,20-dione (progesterone) can be obtained from cows' milk during pregnancy.

A method has been developed for the estimation of 17- and 20-oxogenic steroids in milk. It consists of an initial saponification by boiling 50 ml samples of milk diluted with an equal volume of water for 1 h with 10 g potassium hydroxide followed by gentle mechanical extraction with diethyl ether. The non-saponifiable lipids are then oxidized with 7·5% (w/v) CrO₃ in 50% aqueous acetic acid. A Girard separation of the oxidation products is used to obtain a ketonic fraction which is chromatographed using chloroform (A.R.) on thin layers of silica gel. The areas corresponding to steroid ketones are eluted and the extracts subjected to gas chromatography on a QF-1 coated column.

In addition to measuring progesterone, this procedure would convert