Add time:08/05/2019         Source:sciencedirect.com

17α-estradiol 17-N-acetylglucosaminide (17αE2 17NAG) is an estrogen metabolite hitherto obtained only in rabbits. To gain insight into this unique conjugate, an enzyme immunoassay (EIA) was established by using antiserum elicited against 3-[3-(1-carboxypropyl)] ether of 17αE2 17NAG-bovine serum albumin conjugate; horseradish peroxidase, as a label; and 3,3′,5,5′-tetramethylbenzidine, as a chromogen. The method proved to be specific, and the detection range of the assay was 0.20–10.00 ng/ml. A proposed double conjugate, 3-glucuronide of 17αE2 17NAG, was synthesized to validate the EIA. The EIA was applied to the determination of the urinary level of 17αE2 17NAG in male and female (pregnant and non-pregnant) rabbits with and without β-glucuronidase-sulfatase preparation from Helix pomatia. The results showed that 17αE2 17NAG was mainly excreted as a double conjugate (17αE2 17NAG 3-glucuronide and/or 3-sulfate) and that its level varies during pregnancy.

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