Reproducibility and Robustness of the Xevo™ G3 QTof Mass Spectrometer for the Analysis of Large Cohort Studies

Significance of the Topic

Large-scale metabolomic studies demand highly robust and reproducible LC-MS systems to minimize batch effects and ensure reliable biomarker detection across extensive sample cohorts.

Study Objectives and Overview

The study evaluates the reproducibility and robustness of the Waters Xevo G3 QTof mass spectrometer coupled with an ACQUITY Premier UPLC for analyzing 1,000 injections of human urine over seven days.

Methodology

• Sample preparation: Pooled human urine (NIST SRM 3671) spiked with Waters LCMS QC Reference Standard.
• Chromatography: ACQUITY Premier HSS T3 column (100 × 2.1 mm, 1.8 µm) at 45 °C, 1 µL injection, 0.6 mL/min flow, 10 min gradient.
• Mass spectrometry: Xevo G3 QTof, positive ion mode, MSE continuum acquisition (m/z 50–1200) at 10 Hz, StepWave transmission.

Instrumentation Used

• ACQUITY Premier FTN system with UPLC HSS T3 column
• Xevo G3 QTof mass spectrometer controlled by waters_connect software
• Data processing with MARS, Skyline Daily, and Microsoft Excel

Key Results and Discussion

• Retention time stability: standard deviation 0.084–0.282 s (average ±0.12 s).
• Mass accuracy: average error ±1.2 ppm with SD ±0.79 ppm across 1,000 injections.
• Peak area reproducibility: relative standard deviation 3.8–12.6% (average 6.9%).
• Performance monitored for eight compounds (five spiked standards and three endogenous metabolites).

Benefits and Practical Applications

• Reduced analytical drift and batch effects during extended runs.
• Enhanced confidence in quantitative metabolomic profiling for large cohort studies.
• Fewer QC failures and consistent data quality across long-term analyses.

Future Trends and Applications

• Adoption of real-time quality control and AI-driven drift correction.
• High-throughput multiplexing and extended dynamic range workflows.
• Expansion to diverse biofluid matrices and integration with other omics platforms.

Conclusion

The Xevo G3 QTof combined with the ACQUITY Premier system delivers exceptional reproducibility and robustness for large cohort metabolomic analyses, ensuring stable retention times, accurate mass measurements, and consistent peak responses over extended operation.

References

• Improved Transmission of Labile Species on the Xevo G3 QTof Mass Spectrometer with the StepWave XS, Waters Application Brief 720007794, November 2022.