Illustration of the Analytical Performance of the Waters MassTrak Endocrine Steroid Calibrators and QC Set Using the ACQUITY UPLC I-Class and Xevo TQ-S micro Mass Spectrometer

Significance of the Topic

The accurate quantification of steroid hormones is essential in clinical diagnostics, endocrinology research, and therapeutic monitoring. Reliable calibrators and quality controls underpin consistent results across laboratories, enabling precise measurement of analytes such as cortisol, testosterone, and progesterone.

Objectives and Study Overview

This application note examines the analytical performance of the MassTrak Endocrine Steroid Calibrator and QC Set (IVD) when used with the Waters ACQUITY UPLC I-Class and Xevo TQ-S micro IVD System. The primary goals are to assess sensitivity, selectivity, linearity, precision, and accuracy for a panel of 12 steroid hormones in human serum.

Methodology and Instrumentation

Sample preparation involved protein precipitation and cleanup using Oasis MAX µElution plates. Key steps:

• Mix 125 µL serum with internal standards and methanol, dilute, centrifuge.
• Apply supernatant to MAX plate; wash and elute into collection plate.

Chromatographic separation was achieved on a CORTECS C8 column (2.1 × 100 mm, 2.6 µm) with a gradient of 0.1 mM ammonium fluoride in water (A) and methanol (B) at 0.3 mL/min. Mass spectrometry employed ESI in positive and negative polarity with MRM acquisition on the Xevo TQ-S micro. Data were controlled by MassLynx IVD Software v4.2 and processed via TargetLynx XS.

Main Results and Discussion

Performance metrics for the 12 steroids demonstrated:

• Calibration linearity: r2 values > 0.99 across physiological ranges.
• Sensitivity: Signal-to-noise ratios sufficiently high for low-level detection.
• Precision: Total CVs and repeatability < 10% over five runs (n = 25).
• Accuracy: Mean bias within ± 15% compared to external quality assessment values.

These figures illustrate robust quantification with low variability and strong correlation to reference methods.

Benefits and Practical Applications

The combined calibrator/QC set and UPLC-MS/MS system offers:

• Traceable, lyophilized serum calibrators for consistent method standardization.
• High throughput analysis of multiple steroids in a single run.
• Enhanced assay sensitivity and specificity for clinical laboratories.

Future Trends and Opportunities

Potential developments include:

• Automation of sample preparation workflows to increase throughput.
• Expansion to additional endocrine biomarkers and alternative matrices (e.g., dried blood spots).
• Integration with laboratory information systems and AI-driven data analysis for real-time quality control.

Conclusion

The Waters MassTrak Endocrine Steroid Calibrator and QC Set, paired with the ACQUITY UPLC I-Class and Xevo TQ-S micro IVD System, delivers sensitive, selective, and reproducible quantification of key steroid hormones. This platform supports rigorous clinical testing and research applications with demonstrated precision and accuracy.