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Journal of Dairy Science Vol. 48 No. 7 993-994
© 1965 by American Dairy Science Association ®
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Comparison of Methods for Hydrolysis of Bovine Urinary Estrogen Conjugates1

W. R. Gomes3, T. N. Mellin4 and R. E. Erb

Department of Animal Sciences Purdue University Lafayette, Indiana

ABSTRACT

The accurate measurement of urinary estrogens2 depends on the quantitative hydrolysis of conjugated compounds. As reviewed by Mellin et al. (4) alkaline hydrolysis only partially cleaves estrogen conjugates, and both estrone and estradiol are partially destroyed. Solvent hydrolysis apparently cleaves only sulfate conjugates. Acid hydrolysis completely cleaves conjugated estrogens but destroys a majority of the 17 {alpha}-estradiol present (3, 5, 6). Enzyme hydrolysis appears nondestructive, but incompletely hydrolyzes a mixture of conjugates and may have reduced effectiveness in the presence of certain inhibitors (1). Mellin (3) found that beef liver ß-glucuronidase yielded slightly greater levels of free estrogens than did bacterial ß-glucuronidase when both were incubated with bovine urine.

The purpose of this study was to compare acid hydrolysis, enzyme hydrolysis, and an enzyme-acid hydrolysis sequence for effectiveness in cleaving conjugates of 14C-labeled estrogens in bovine urine.

EXPERIMENTAL PROCEDURE

Thirty-six microcuries of 17 ß-estradiol-4-14C (New England Nuclear Corp.) were administered intravenously to a nonpregnant yearling heifer in eight injections (4.5 µc/injection).

FOOTNOTES

1 Journal Paper no. 2550, Purdue University Agricultural Experiment Station, Project 1306.

3 Present address: Department of Dairy Science, Ohio State University, Columbus, Ohio.

4 Present address: Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts.






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Copyright © 1965 by the American Dairy Science Association ®.