Open AccessSimultaneous quantitation of four androgens and 17‐hydroxyprogesterone in polycystic ovarian syndrome patients by LC‐MS/MS
Author(s) -
Cao Zheng,
Lu Yifan,
Cong Yuting,
Liu Ying,
Li Youran,
Wang Husheng,
Zhang Qiaoli,
Huang Wenxi,
Liu Jingrui,
Dong Ying,
Tang Guodong,
Luo Yiqi R.,
Yin Chenghong,
Zhai Yanhong
Publication year - 2020
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.23539
Subject(s) - chromatography , chemistry , polycystic ovary , analyte , electrospray ionization , selected reaction monitoring , liquid chromatography–mass spectrometry , androstenedione , androgen , protein precipitation , endocrinology , testosterone (patch) , medicine , tandem mass spectrometry , mass spectrometry , hormone , insulin , biochemistry , insulin resistance
Background Due to the low concentration of androgens in women and the limitation of immunoassays, it remains a challenge to accurately determine the levels of serum androgens in polycystic ovary syndrome (PCOS) patients for clinical laboratories. In this report, a liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed and validated for simultaneous quantitation of testosterone (T), androstenedione (A4), dehydroepiandrosterone sulfate (DHEAS), dihydrotestosterone (DHT), and 17‐hydroxyprogesterone (17‐OHP) that are associated with PCOS. Methods The serum samples were processed by protein precipitation and solid phase extraction before analysis with the in‐house developed LC‐MS/MS. The chromatographic separation was achieved with a C18 column, using a linear gradient elution with two mobile phases: 0.02% formic acid in water (phase A) and 0.1% formic acid in methanol (phase B). The separated analytes were detected by positive or negative electrospray ionization mode under multiple reaction monitoring (MRM). Results The assay for all the five analytes was linear, stable, with imprecision less than 9% and recoveries within ±10%. The lower limits of quantification were 0.05, 0.05, 5, 0.025, and 0.025 ng/mL for T, A4, DHEAS, DHT, and 17‐OHP, respectively. In the receiver operating characteristic curve (ROC) analyses with the PCOS (n = 63) and healthy (n = 161) subjects, the AUC of the four‐androgen combined was greater than that of any single androgen tested in PCOS diagnosis. Conclusions The LC‐MS/MS method for the four androgens and 17‐OHP showed good performance for clinical implementation. More importantly, simultaneous quantitation of the four androgens provided better diagnostic power for PCOS.