LC/MS, LC/MS/MS, LC/QQQ
IndustriesClinical Research
ManufacturerWaters
Importance of the Topic
Accurate measurement of steroid hormones in human serum is essential for diagnosing endocrine disorders, monitoring therapy efficacy, and conducting clinical research. The ACQUITY UPLC I-Class/Xevo TQ-S micro IVD System offers a highly sensitive and selective platform for quantifying key androgens, progestogens, and glucocorticoids with rapid turnaround and minimal sample volume.
Objectives and Study Overview
The study aimed to evaluate the analytical performance of the ACQUITY UPLC I-Class/Xevo TQ-S micro IVD System for seven steroids in human serum: testosterone, androstenedione, 17-hydroxyprogesterone, dehydroepiandrosterone sulfate (DHEAS), cortisol, 11-deoxycortisol, and 21-deoxycortisol. Key performance parameters included calibration linearity, limit of quantification, precision, accuracy, and external quality assessment bias.
Methodology and Instrumentation
Sample Preparation:
- 100 µL serum precipitated with methanol, diluted with water, and centrifuged.
- Solid-phase extraction using Oasis PRiME HLB µElution plates.
Liquid Chromatography Conditions:
- Column: ACQUITY UPLC HSS T3 (1.8 µm, 2.1 × 50 mm) with VanGuard HSS T3 pre-column (1.8 µm, 2.1 × 5 mm).
- Mobile phases: 2 mM ammonium acetate + 0.1% formic acid in water (A) and in methanol (B).
- Flow rate: 0.6 mL/min; gradient from 45% to 98% B over 4 min total.
Mass Spectrometry Conditions:
- Instrument: Xevo TQ-S micro IVD; ESI positive/negative.
- Acquisition: MRM with 0.75 FWHM resolution in both MS1 and MS2.
Main Results and Discussion
The system achieved baseline chromatographic selectivity for all analytes. Calibration curves were linear (r²>0.99) across concentration ranges of 0.09–43 000 nmol/L. Limits of quantification ranged from 0.09 to 65 nmol/L with precision at LLOQ between 4.0% and 14.1% RSD. Total precision and repeatability for quality controls were better than 7.6% and 5.2% RSD, respectively. External quality assessment biases remained within ±5.8%. These results demonstrate high sensitivity, reproducibility, and accuracy suitable for clinical diagnostics.
Benefits and Practical Applications
- Minimal sample volume (100 µL) and streamlined preparation support high throughput.
- Robust method with short run times accelerates laboratory workflow.
- Comprehensive steroid panel meets diverse clinical testing needs.
Future Trends and Applications
Expansion of the method to additional steroid metabolites and related biomarkers could further enhance diagnostic panels. Integration with automated sample handling and data processing will improve throughput. Emerging applications include personalized medicine, real-time therapeutic monitoring, and adaptation for point-of-care platforms.
Conclusion
The ACQUITY UPLC I-Class/Xevo TQ-S micro IVD System provides sensitive, selective, and precise quantification of key steroid hormones in serum. Its performance characteristics make it well suited for routine clinical and research laboratories focused on endocrine analysis.
Reference
Waters Corporation. ACQUITY UPLC I-Class/Xevo TQ-S micro IVD System: Analytical Performance for Androgens, Progestogens and Glucocorticoids. August 2018.
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