Simultaneous profiling of classic and 11-oxygenated androgens via a novel derivatization method coupled with HPLC-MS/MS: A game-changer for endocrine disorders
Xianhua Zhang,
Huiyu Xu,
Yue Sun,
Xin Xiong,
Lingling Liu,
Li Yang,
Libo Zhao,
Xiaona Li,
Rong Li,
Rongsheng Zhao
Affiliations
Xianhua Zhang
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Huiyu Xu
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, PR China; Key Laboratory of Assisted Reproduction, Peking University, Ministry of Education, Beijing 100191, PR China; National Clinical Research Center for Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, PR China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing 100191, PR China
Yue Sun
Tianjin Wego Medical Technology Co., LTD, Tianjin 300450, PR China
Xin Xiong
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Lingling Liu
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Li Yang
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Libo Zhao
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Xiaona Li
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China
Rong Li
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, PR China; Key Laboratory of Assisted Reproduction, Peking University, Ministry of Education, Beijing 100191, PR China; National Clinical Research Center for Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, PR China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing 100191, PR China; Corresponding authors.
Rongsheng Zhao
Department of Pharmacy, Peking University Third Hospital, Beijing 100191, PR China; Therapeutic Drug Monitoring and Clinical Toxicology Center for Peking University, Beijing 100191, PR China; NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Beijing 102206, PR China; Corresponding authors.
The accumulation of classic or 11-oxygenated androgens is a hallmark of polycystic ovary syndrome (PCOS). Traditional electrochemiluminescence immunoassays (ECLIA) are limited in their ability to measure a broad spectrum of androgens and may suffer from potential cross-reactivity, leading to inaccurate results. In this study, we developed a highly sensitive HPLC-MS/MS method for the simultaneous determination of nine androgens, including four classic and five 11-oxygenated androgens in human serum. Magnetic solid-phase extraction (MSPE) using magnetic graphene oxide nanoparticles followed by in-situ derivatization further improved the detection efficiency. A novel quaternary ammonium aminooxy (QAO) reagent was employed as the derivatization agent, featuring a permanently charged MS-tag and an aminooxy group that reacts with steroid ketone groups. This derivatization significantly enhanced MS sensitivity compared to traditional derivatization agents. The linear ranges for different targets were set based on clinical use and the lower limit of quantification (LLOQ) of the target androgens ranged from 0.005 to 1 ng/mL. The calibration curves exhibited excellent linear correlation (r ≥ 0.99) across the validated ranges for all the analytes. The intra- and inter-run precision was below 9.8 % and 9.7 % for the lower limit of quantification (LLOQ), and 7.5 % and 6.9 % for quality controls (QCs). Intra- and inter-day relative errors for LLOQ ranged from -8.2 % to 10.7 %, and from -7.4 % to 13.8 % for QCs. Extraction recoveries varied from 41.8 % to 80.2 %. The method was applied to 107 clinical samples, demonstrating it as a fast, sensitive and accurate method for androgens’ quantification. By offering the ability to detect and quantify a broader range of androgens, including those not measurable by ECLIA, and avoiding potential cross-reactions. This method provides a superior analytical tool for the diagnosis and management of PCOS in clinical settings.
