Simultaneous quantitation of four androgens and 17‐hydroxyprogesterone in polycystic ovarian syndrome patients by LC‐MS/MS. Issue 12 (21st August 2020)

• Record Type: Journal Article
• Authors: 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
• Is Part Of: Journal of clinical laboratory analysis. Volume 34:Issue 12(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-08-21
• Subjects: androgen -- diagnosis -- LC‐MS/MS -- PCOS -- validation; Diagnosis, Laboratory -- Periodicals; Medical laboratory technology -- Periodicals; 616
• DOI: 10.1002/jcla.23539
• Languages: English
• ISSNs: 0887-8013

Abstract

Abstract: 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. Abstract : An in‐house 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 polycystic ovary syndrome (PCOS). With PCOS patients and healthy controls, the combinational detection of the four androgens (T, A4, DHEAS, DHT) was superior to any of the single androgen in PCOS diagnosis. (A: The complete chromatograms of the five analytes resolved by the established LC‐MS/MS method: DHEAS (1), A4 (2), T (3), 17‐OHP (4), and DHT (5). B: The ROC analyses were performed with the 63 PCOS patients and 161 healthy controls. The combination detection referred to the simultaneous measurements of the four androgens of T, A4, DHEAS, DHT).