Clinical Usefulness of Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Low Serum Testosterone Measurement
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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.
Abstract
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.
참고문헌 (27)
- Zhou H, "Simultaneous measurement of total estradiol and testosterone in human serum by isotope dilution liquid chromatography tandem mass spectrometry", Anal Bioanal Chem, 409, pp.5943~5954 (2017)
- Rosner W, "Position statement: utility, limitations, and pitfalls in measuring testosterone: an endocrine society position statement", J Clin Endocrinol Metab, 92, pp.405~413 (2007)
- Zhou Y, "A robust LC-MS", MS assay with online cleanup for measurement of serum testosterone, 2019, pp.42, 2561~2568
- Herold DA, "Immunoassays for testosterone in women : better than a guess?", Clin Chem, 49, pp.1250~1251 (2003)
- Rosner W, "Toward excellence in testosterone testing: a consensus statement", J Clin Endocrinol Metab, 95, pp.4542~4548 (2010)
- La’ulu SL, "How low can you go? Analytical performance of five automated testosterone immunoassays", Clin Biochem, 58, pp.64~71 (2018)
- Vesper HW, "CDC project on standardizing steroid hormone measurements", Steroids, 73, pp.1286~1292 (2008)
- "Centers for Disease Control and Prevention, Hormone Standardization Program (CDC HoSt)" (2021)[URL]
- "Bioanalytical method validation guidance for industry" (2018)[URL]
- CLSI, "Liquid chromatography-mass spectrometry methods. 1st ed. C62-A", Clinical and Laboratory Standards Institute, ~ (2014)
- CLSI, "Evaluation of linearity of quantitative measurement procedures. 2nd ed. EP06-ED2", Clinical and Laboratory Standards Institute, ~ (2020)
- French D, "Development and validation of a serum total testosterone liquid chromatography-tandem mass spectrometry(LC-MS", MS)assay calibrated to NIST SRM 971, 2013, pp.415, 109~117
- Rappold Brian A., "Review of the Use of Liquid Chromatography-Tandem Mass Spectrometry in Clinical Laboratories: Part I-Development", Annals of Laboratory Medicine, 42(2), pp.121~140 (2022), 대한진단검사의학회[DOI]
- Rappold Brian A., "Review of the Use of Liquid Chromatography-Tandem Mass Spectrometry in Clinical Laboratories: Part II–Operations", Annals of Laboratory Medicine, 42(5), pp.531~557 (2022), 대한진단검사의학회[DOI]
- Yun YM, "Performance criteria for testosterone measurements based on biological variation in adult males : recommendations from the Partnership for the Accurate Testing of Hormones", Clin Chem, 58, pp.1703~1710 (2012)
- CLSI, "Defining, establishing, and verifying reference intervals in the clinical laboratory" (2010), Clinical and Laboratory Standards Institute
- Moal V, "Low serum testosterone assayed by liquid chromatography-tandem mass spectrometry. Comparison with five immunoassay techniques", Clin Chim Acta, 386, pp.12~19 (2007)
- Kushnir MM, "Performance characteristics of a novel tandem mass spectrometry assay for serum testosterone", Clin Chem, 52, pp.120~128 (2006)
- McCartney CR, "Estimating testosterone concentrations in adolescent girls : comparison of two direct immunoassays to liquid chromatography-tandem mass spectrometry", Steroids, 140, pp.62~69 (2018)
- Vesper HW, "Interlaboratory comparison study of serum total testosterone measurements performed by mass spectrometry methods", Steroids, 74, pp.498~503 (2009)
- Cao ZT, "Accuracy-based proficiency testing for testosterone measurements with immunoassays and liquid chromatography-mass spectrometry", Clin Chim Acta, 469, pp.31~36 (2017)
- 최리화, "Dried Blood Spot Multiplexed Steroid Profiling Using Liquid Chromatography Tandem Mass Spectrometry in Korean Neonates", Annals of Laboratory Medicine, 39(3), pp.263~270 (2019), 대한진단검사의학회[DOI]
- Ko DH, "Simultaneous measurement of serum chemical castration agents and testosterone levels using ultra-performance liquid chromatography-tandem mass spectrometry", J Anal Toxicol, 40, pp.294~303 (2016)
- Olisov D, "Measurement of serum steroid profiles by HPLC-tandem mass spectrometry", J Chromatogr B Analyt Technol Biomed Life Sci, 1117, pp.1~9 (2019)