Enzyme immunoassay for the measurement of 17alpha-estradiol 17-N-acetylglucosaminide in rabbit urine.
17alpha-estradiol 17-N-acetylglucosaminide (17alphaE2 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 17alphaE2 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 17alphaE2 17NAG, was synthesized to validate the EIA. The EIA was applied to the determination of the urinary level of 17alphaE2 17NAG in male and female (pregnant and non-pregnant) rabbits with and without beta-glucuronidase-sulfatase preparation from Helix pomatia. The results showed that 17alphaE2 17NAG was mainly excreted as a double conjugate (17alphaE2 17NAG 3-glucuronide and/or 3-sulfate) and that its level varies during pregnancy.
Hatano Research Institute, Food and Drug Safety Center, Hadano, Kanagawa, Japan. madokan@qc4.so-net.ne.jp
M
M
Nakagomi
K
K
Yamada
Y
Y
Matsuki
H
H
Kurihara
E
E
Suzuki
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000652:urine; D000818:Animals; D004958:Estradiol / Q000031:analogs & derivatives* / Q000652:urine; D005260:Female; D005966:Glucuronidase / Q000378:metabolism; D006868:Hydrolysis; D007124:Immunoenzyme Techniques; D008297:Male; D011247:Pregnancy; D011817:Rabbits; D012680:Sensitivity and Specificity; D013429:Sulfatases / Q000378:metabolism
C121106:estradiol 17-acetylglucosaminide; D004958:Estradiol; D013429:Sulfatases; D005966:Glucuronidase; D000117:Acetylglucosamine