1. Profiling endogenous serum estrogen and estrogen-glucuronides by liquid chromatography--tandem mass spectrometry
- Author
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Caron, Patrick, Audet-Walsh, Etienne, Lepine, Johanie, Belanger, Alain, and Guillemette, Chantal
- Subjects
Estrogen -- Measurement ,Estrogen -- Research ,Liquid chromatography -- Usage ,Mass spectrometry -- Usage ,Chemistry - Abstract
Estrogens, namely, 17[beta]-estradiol ([E.sub.2]), are conjugated to glucuronides (G), and this metabolic conversion is part of their tissular-concentration control-mechanism. This inactivation process has been observed, in addition to the liver, in several estrogen-dependent tissues and the resulting polar metabolites are detected in circulation. We developed and validated a highly sensitive and specific mass spectrometry-based method to directly measure estrogen-G serum levels. The method uses deuterated standards but does not involve enzymatic hydrolysis, a major improvement over previous techniques. Estrone ([E.sub.1]), [E.sub.1]-sulfate, [E.sub.2], the 3-G of [E.sub.1], [E.sub.2], 2-methoxy-[E.sub.1] (2-MeO[E.sub.1]) and 2-methoxy-[E.sub.2] (2-MeO[E.sub.2]), and the 17-G of [E.sub.2] were measured in serum of 19 premenopausal and 10 postmenopausal healthy women. Two extractions, solid-phase and liquid--liquid, were performed to isolate the estrogens. Estrogens were then quantified by mass spectrometry in the negative MRM ion mode using an API3200 spectrometer with a turbo ionspray source. The method selectively measured estrogen glucuronides with sensitivity [greater than or equal to] 5 pg/mL accuracy 90-111%, and reproducibility (CV = 1.4-13.3%). The method is applicable between 5 and 1000 pg/mL. For the ovarian follicular phase, the major metabolite found was [E.sub.1]-3G, with [E.sub.2]-3G and 2-MeO[E.sub.1]-3G found in lesser amounts (54, 10.4, and 7.8 pg/mL, respectively) These concentrations are 2.6- to 3-fold greater than found for luteal-phase estrogens. The concentrations of [E.sub.2]-17G and 2-MeO[E.sub.2]-3G were usually less than the limit of quantification. In serum of postmenopaussl women, [E.sub.1]-3G was the most abundant estrogen found (30.9 pg/mL). Our method profiles estrogens and estrogen-glucuronides and may represent a new tool to identify biomarkers in hormone-dependent diseases. 10.1021/ac9019126
- Published
- 2009