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Journal of Dairy Science Vol. 44 No. 2 300-309
© 1961 by American Dairy Science Association ®
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Quantitative Methods for Determining Progestins in Corpora Lutea, Ovaries, and Adrenals of the Cow and Sow Using Progesterone-4-C14 as a Measure of Recovery of Extracted Hormone1

F. Stormshak2, 3,, Marjorie L. Hunt and R. E. Erb

Departments of Dairy Science and Agricultural Chemistry, Washington State University, Pullman

ABSTRACT

A quantitative method is described for the chemical assay of progesterone and {Delta}4-pregnene-20ß-ol-3-one. The tissues were extracted with ethyl acetate, followed by vacuum distillation and partitioning of the residue between Skellysolve B - 70% methanol and benzene. The progestins were separated and further purified by paper chromatography using the Bush Skellysolve B - 95% methanol system, eluted, and rechromatographed in the same system. Progesterone-4-C14 (0.08 µg; 3,500 c.p.m.) was added at the time of initial extraction to estimate recovery of progesterone during each assay. The radiochemical purity of the compound was 93–94%. About 4% of the radioactivity remained in the tissue during initial extraction and approximately 16% was lost during each elution of the paper chromatograms. Average recovery of progesterone-4-C14 was 40% for 224 samples of cow tissues (standard error 0.7%) and 51% for 114 samples of sow tissues (standard error 1.1%). No {Delta}4-pregnene-20ß-ol-3-one was detected in sow tissues.

FOOTNOTES

1 Scientific Paper No. 2022, Washington Agricultural Experiment Stations. Project 1395-195. This investigation was supported in part by funds for biological and medical research by the State of Washington, Initiative Measure No. 171.

2 The data are from a thesis submitted by the senior author to the graduate faculty, Washington State University, in partial fulfillment of the requirements for the degree of Master of Science, August, 1960.

3 Present address: Department of Genetics, University of Wisconsin, Madison.






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Copyright © 1961 by the American Dairy Science Association ®.