A quantitative analysis of total and free 11-oxygenated androgens and its application to human serum and plasma specimens using liquid-chromatography tandem mass spectrometry

被引：24

作者：

Caron, Patrick ^{[1
]}

Turcotte, Veronique ^{[1
]}

Guillemette, Chantal ^{[1
,2
]}

机构：

[1] Univ Laval, Univ Laval Res Ctr, CHU Quebec, Pharmacogen Lab, Quebec City, PQ, Canada

[2] Canada Res Chair Pharmacogen, Quebec City, PQ, Canada

来源：

JOURNAL OF CHROMATOGRAPHY A | 2021年 / 1650卷

基金：

加拿大健康研究院;

关键词：

11-oxygenated androgens; Adrenal androgens; Mass spectrometry; Validation; ADRENAL 11-BETA-HYDROXYANDROSTENEDIONE METABOLITES; PREDOMINANT ANDROGENS; 19-CARBON STEROIDS; C19; STEROIDS; LC-MS/MS; 11-KETOTESTOSTERONE; ANDROSTENEDIONE; QUANTIFICATION; BIOSYNTHESIS; CELLS;

D O I：

10.1016/j.chroma.2021.462228

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Bioactive 11-oxygenated C 19 adrenal-derived steroids (11-oxy C 19 ) are potentially relevant in diverse endocrine and metabolic contexts. We report the development and validation of a liquid chromatog-raphy electrospray ionization tandem mass spectrometric method (LC-ESI-MS/MS) for the simultane-ous quantification of seven 11-oxy C 19 using 200 mu L of plasma or serum. Sample preparation in-volved chemical derivatization using hydroxylamine after liquid-liquid extraction to improve speci-ficity and sensitivity. The method allowed the quantitation of total 11-oxy C 19 (free + sulfate and glucuronide conjugates) following enzymatic hydrolysis. This included the abundant precursor 11-hydroxyandrostenedione (11OHA4) and the most potent androgenic derivatives 11-keto-testosterone (11KT) and 11-keto-dihydrotestosterone (11KDHT), their abundant metabolites 11-hydroxyandrosterone (11OHAST) and 11-keto-androsterone (11KAST) potentially feeding back into the pool of potent andro -gens, in addition to 11-keto-androstenedione (11KA4) and 11-hydroxytestosterone (11OHT). Stable iso-topes were used as internal standards, and calibrators and quality controls were prepared in the same matrix as the study samples. Performance was validated against the Food and Drug Administration Cri -teria. The method was sensitive with lower limit of quantification (LLOQ) values of 10 and 20 pg/mL for free and total 11-oxy C 19 , respectively. The applicability was demonstrated in men and women adult donors that showed sex-differences. All steroids were quantified well above LLOQ, except 11KDHT that remained undetectable suggesting interfering endogenous molecules present in non-derivatized samples in which a peak was observed. By providing accurate and reliable quantitative data, this method will permit to evaluate how profiling of 11-oxy C 19 will be most informative as diagnostic, prognostic and/or theranostic tools. (c) 2021 Elsevier B.V. All rights reserved.

引用

页数：12

共 45 条

[1] Profiling of Endogenous Estrogens, Their Precursors, and Metabolites in Endometrial Cancer Patients: Association with Risk and Relationship to Clinical Characteristics [J].