Clinical Usefulness of Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Low Serum Testosterone Measurement

Cho Sung-Eun; Han Jungsun; Park Ju-Hee; Park Euna; Kim Geun Young; Lee Jun Hyung; Yi Ahram; Lee Sang Gon; Lee Eun Hee; Yun Yeo-Min

doi:10.3343/alm.2023.43.1.19

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자료유형: 학술저널

저자정보: Cho Sung-Eun (Department of Laboratory Medicine GCLabs Yongin Korea) Han Jungsun (Department of Laboratory Medicine GCLabs Yongin Korea) Park Ju-Hee (Department of Laboratory Medicine GCLabs Yongin Korea) Park Euna (Department of Laboratory Medicine GCLabs Yongin Korea) Kim Geun Young (Department of Laboratory Medicine GCLabs Yongin Korea) Lee Jun Hyung (Department of Laboratory Medicine GCLabs Yongin Korea) Yi Ahram (Department of Laboratory Medicine GCLabs Yongin Korea) Lee Sang Gon (Department of Laboratory Medicine GCLabs Yongin Korea) Lee Eun Hee (Department of Laboratory Medicine GCLabs Yongin Korea) Yun Yeo-Min (Department of Laboratory Medicine Konkuk University Medical Center Konkuk University School of Medicine Seoul Korea)

저널정보: 대한진단검사의학회 Annals of Laboratory Medicine Annals of Laboratory Medicine 제43권 제1호

발행연도: 2023.1

수록면: 19 - 28 (10page)

DOI: 10.3343/alm.2023.43.1.19

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Background: Mass spectrometry methods exhibit higher accuracy and lower variability than immunoassays at low testosterone concentrations. We developed and validated an ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay for quantifying serum total testosterone. Methods: We used an ExionLC UPLC (Sciex, Framingham, MA, USA) system and a Sciex Triple Quad 6500+ (Sciex) MS/MS system in electrospray ionization and positive ion modes with multiple reaction monitoring transitions to evaluate precision, accuracy, linearity, lower limit of quantitation (LLOQ), carryover, ion suppression, stability, and reference intervals. For method comparison, we measured serum testosterone concentrations using this method in 40 subjects whose testosterone concentrations ranged from 0.14 to 55.48 nmol/L as determined using the Architect i2000 immunoassay (Abbott Diagnostics, Abbott Park, IL, USA) and in an additional 160 sera with testosterone concentrations <1.67 nmol/L. Results: The intra- and inter-run precision CVs were <2.81%, and the accuracy bias values were <3.85%, which were all acceptable. The verified linear interval was 0.03–180.84 nmol/L; the LLOQ was 0.03 nmol/L. No significant carryover and ion suppression were observed. The testosterone in serum was stable at 4°C, at –20°C, and after three freeze-thaw cycles. The reference intervals were successfully verified. The correlation was good at testosterone concentrations of 0.14–55.48 nmol/L; however, the Architect assay showed positive percent bias at concentrations <1.67 nmol/L. Conclusions: The UPLC-MS/MS assay shows acceptable performance, with a lower LLOQ than the immunoassay. This method will enable the quantitation of low testosterone concentrations.

#Tandem mass spectrometry #Testosterone #Quantitation #Performance

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