4.4 Article

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

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ANNALS OF LABORATORY MEDICINE
卷 43, 期 1, 页码 19-+

出版社

KOREAN SOC LABORATORY MEDICINE
DOI: 10.3343/alm.2023.43.1.19

关键词

Tandem mass spectrometry; Testosterone; Quantitation; Performance

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Ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay shows acceptable performance and a lower limit of quantitation (LLOQ) compared to immunoassays for quantifying serum total testosterone, enabling the quantitation of low testosterone concentrations.
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 degrees C, at -20 degrees 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.

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