Defining hyperandrogenism in polycystic ovary syndrome: measurement of testosterone and androstenedione by liquid chromatography–tandem mass spectrometry and analysis by receiver operator characteristic plots

Abstract

Objective: Hyperandrogenism is one of the diagnostic criteria for the polycystic ovary syndrome (PCOS) despite no agreed definition of hyperandrogenism. In part, this is due to the quality of testosterone immunoassays. We have developed liquid chromatography-tandem mass spectrometry methods for analysing testosterone and androstenedione (Ad) to study their reference ranges and diagnostic utility in PCOS. Design, setting and subjects: A consecutive series of 122 women attending a reproductive medicine clinic. Methods: Blood samples were taken during the early follicular phase for measurement of LH, FSH, oestradiol, Ad, testosterone and sex hormone-binding globulin (SHBG). Retrospective case note analysis was used to determine the clinical features and ultrasound findings. Results: The incidence of PCOS was 13.9%. The reference interval for testosterone was !1.8 nmol/l and for Ad was 1.4-7.4 nmol/l. There were significant differences in total testosterone (PZ0.001), Ad (P!0.05) and free androgen index (FAI; P!0.0001) between the women with and without PCOS. Diagnostic performance using receiver operator characteristic plots showed area under the curve (AUC) for FAI 0.81, testosterone 0.75 and Ad 0.66. The AUC for the LH:FSH ratio was 0.72. Conclusions: Our analysis of a consecutive series of women attending a reproductive clinic has provided an appropriate series on which to construct reference ranges for key androgens in women. Secondly, it has allowed us to conclude that early follicular serum testosterone measured using tandem mass spectrometry, FAI and the LH:FSH ratio are valuable laboratory tests in the diagnosis of PCOS.

Assessment of androgen concentration in women: liquid chromatography–tandem mass spectrometry and extraction RIA show comparable results

“…Androgen concentrations for women with PCOS were significantly higher compared with female controls and women affected by POI, which confirms other recent studies applying LC-MS/MS (26,27). However, this is the first study to measure androgens with LC-MS/MS in women with POI.…”

“…Intraclass correlation coefficients (ICC) with 95% CIs were calculated for testosterone, DHEA, and AD. Large ICC (maximum 1) indicates that variation between both assays is equal to zero (perfect agreement) (26,27). Because androgen concentrations were not normally distributed, logarithmic transformation of variables was applied before execution of analyses.…”

Sex hormone measurements using mass spectrometry and sensitive extraction radioimmunoassay and risk of estrogen receptor negative and positive breast cancer: Case control study in UK Collaborative Cancer Trial of Ovarian Cancer Screening (UKCTOCS)

“…Estradiol by extraction radioimmunoassay Unconjugated estradiol was measured in 100 uL of sample in duplicate using an in-house radioimmunoassay (25). Estradiol was extracted into 1 mL diethyl ether (Vickers) by vortex mixing for 3 min (after addition of 10 uL 2M sodium hydroxide).…”

Markers of hyperandrogenism in South Asians with polycystic ovary syndrome

“…The incidence of PCOS being 13.9%, the reference interval for testosterone was 1.8 nmol/l with the conclusion that early follicular phase serum testosterone measured using tandem mass spectrometry and FAI are valuable in the laboratory diagnosis of PCOS (13).…”