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Updated: Nov 16, 2025
Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry UPLC-MS
Published on: March 14, 2013
Automated Sample Preparation Enables LC-MS/MS as a Routine Diagnostic Analysis for Serum Testosterone
Judith A Stone1, Dave R van Staveren2, Robert L Fitzerald1,3
1Toxicology/Mass Spectrometry Laboratory, University of California San Diego Health System Center for Advanced Laboratory Medicine, San Diego, CA.
Automated liquid handling with AC Extraction Plate™ improves LC-MS/MS for serum testosterone testing, offering enhanced accuracy and reproducibility for clinical laboratories. This method simplifies sample preparation, making LC-MS/MS more accessible for routine testosterone analysis.
Area of Science:
- Clinical Chemistry
- Mass Spectrometry
- Endocrinology
Background:
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers superior accuracy and specificity for serum testosterone analysis compared to immunoassays.
- Technical challenges in sample preparation have historically limited LC-MS/MS adoption in clinical laboratories, despite its advantages.
- Improving reproducibility in LC-MS/MS testosterone assays remains a key objective for widespread clinical implementation.
Purpose of the Study:
- To evaluate the replacement of automated immunoassay with LC-MS/MS for all serum testosterone analyses.
- To assess the performance of a novel automated sample preparation method using AC Extraction Plate™ for LC-MS/MS testosterone testing.
- To validate the robustness and clinical applicability of the developed LC-MS/MS method.
Main Methods:
- Implemented LC-MS/MS with automated liquid handling and a novel AC Extraction Plate™ for sample preparation.
- Validated the LC-MS/MS method for key performance parameters including matrix effect, recovery, precision, accuracy, specificity, and reportable range.
- Performed correlation studies with existing testosterone testing methods and evaluated calibration stability.
Main Results:
- The LC-MS/MS method demonstrated excellent performance with a wide reportable range (4-1560 ng/dL) and high accuracy (<4.0% bias vs. CRM).
- Excellent precision was achieved with between-day coefficients of variation (CV) <6%, and no significant interference was observed from common sample components or confounding substances.
- The method showed strong agreement with other LC-MS/MS assays (mean bias 1.3%) and demonstrated feasibility for routine use with sparse/historical calibration.
Conclusions:
- Automated sample extraction using the AC Plate™ facilitates ease of use for LC-MS/MS testosterone testing.
- The developed LC-MS/MS method offers robust performance suitable for adoption in routine clinical laboratory settings.
- This approach addresses previous technical hurdles, enabling broader implementation of accurate LC-MS/MS for serum testosterone analysis.
