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Isolation and identification of androstanediol glucuronide from human plasma
P N Rao1, J E Burdett, P H Moore
1Department of Organic Chemistry, Southwest Foundation for Biomedical Research, San Antonio, TX 78284.
Journal of Steroid Biochemistry
|November 1, 1987
Summary
Dihydrotestosterone (DHT) infusion studies reveal that the primary circulating DHT metabolite in humans is 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide, not the 3-glucuronide form.
Area of Science:
- Endocrinology
- Steroid Metabolism
- Analytical Chemistry
Background:
- Dihydrotestosterone (DHT) is a key androgen.
- Understanding DHT metabolism is crucial for hormonal health.
- Previous studies established steady-state conditions for DHT infusion.
Purpose of the Study:
- To identify and quantify dihydrotestosterone (DHT) glucuronide conjugates in human plasma.
- To determine the predominant circulating form of DHT glucuronide.
Main Methods:
- Infusion of [3H]DHT into human subjects.
- Plasma extraction and separation of unconjugated steroids.
- Selective extraction and purification of steroid glucuronides using XAD-2 chromatography.
- High-performance liquid chromatography (HPLC) for conjugate analysis.
- Derivatization of glucuronides to glucuronide triacetyl methyl ester (GAME) for enhanced detection.
Main Results:
- A novel HPLC system successfully separated 3- and 17-conjugated DHT glucuronides.
- Analysis of pooled plasma samples revealed the exclusive presence of the 17-glucuronide conjugate.
- No significant levels of the 3-glucuronide conjugate were detected.
Conclusions:
- The primary circulating metabolite of dihydrotestosterone in humans is 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide.
- This finding clarifies the metabolic fate of DHT in vivo.
- The 17-glucuronide is the biologically relevant circulating form.