Comparison of Methods for Hydrolysis of Bovine Urinary Estrogen Conjugates

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Abstract
Bovine urine containing radioactive metabolites of intravenously administered 17[beta]-estradiol-4-C14 was subjected to different hydrolysis procedures to determine a satisfactory method for freeing conjugates of estrogens. Acid hydrolysis for 18 or 16 min. yielded, respectively, 21 and 6% of the radioactivity. Incubation with [beta]-glucuronidase (Ketodase or Glusulase) liberated 60% of the radioactivity, and the enzyme-acid combination increased this to 83%. Acid hydrolysis alone gave very low yields of 17[alpha]-estradiol (P < .01), whereas enzyme hydrolysis was inefficient for releasing estrone-C14. The most satisfactory method is to 1st hydrolyze with enzyme followed by acid to avoid destruction of 17[alpha]-estradiol and give maximum yields of extractable estrogens.