Standardized Targeted Metabolomics Using the BIOCRATES MxP Quant 500 Kit on the ACQUITY UPLC I-Class PLUS and Xevo TQ-XS Mass Spectrometer

Significance of the Topic

The development of standardized targeted metabolomics workflows is crucial for reliable quantification of metabolites and lipids across laboratories and studies. By enabling multiplexed analysis in a small sample volume, such approaches accelerate biomarker discovery, disease research, and quality control in pharmaceutical and clinical applications.

Objectives and Study Overview

This study validated the BIOCRATES MxP Quant 500 kit on a Waters ACQUITY UPLC I-Class PLUS system coupled to a Xevo TQ-XS mass spectrometer. The assay targets up to 630 metabolites and lipids from 26 compound classes plus 232 biologically meaningful indicators, all derived from a 10-µL sample in a single experiment.

Methodology

Samples (plasma, serum, tissue, fecal homogenate) were processed on a patented 96-well filter plate. A 30-minute drying step was followed by 1-hour PITC derivatization, another drying step, and extraction with methanolic ammonium acetate. Dual analyses comprised reversed-phase UHPLC-ESI-MS/MS (13 small-molecule classes) and flow-injection MS/MS (hexoses and 12 lipid classes) in MRM mode. Data were normalized and quantified via automated MetIDQ software workflows.

Used Instrumentation

• ACQUITY UPLC I-Class PLUS system with MxP Quant 500 UHPLC column
• Xevo TQ-XS triple quadrupole mass spectrometer
• 96-well filter plates and reagents from the MxP Quant 500 kit
• MassLynx v4.2 software and MetIDQ for data acquisition and analysis

Key Results and Discussion

Analysis times were 7 minutes for each LC-MS/MS run and 4 minutes for each FIA-MS/MS run, yielding a 36-hour throughput for 80 samples on a 96-well plate. Validation in human plasma demonstrated intra- and inter-batch accuracy of 85–115% and precision (CV) <15% for seven-point calibrated analytes, classifying them as quantitative. One-point calibrated compounds met CV <20% and accuracy of 80–120%. No analytes were deemed invalid. Comparison with NIST SRM 1950 showed 85–115% accuracy for amino acids, creatinine, and hexoses. Approximately 520 analytes in plasma were routinely quantified >LOD. The Xevo TQ-XS outperformed the TQ-S with ~20 additional detectable analytes, broader dynamic range, shorter dwell times, and reduced total runtime (~33 hours).

Benefits and Practical Applications of the Method

• Standardized, ready-to-use kit with reagents, hardware, and software for end-to-end workflow
• High throughput with minimal sample volume (10 µL)
• Broad coverage of key metabolic pathways and lipid classes
• Robust performance across sample types (blood, tissue, feces) and species
• Automated data processing ensuring batch-to-batch comparability

Future Trends and Applications

Ongoing developments include expanded metabolite panels, integration with microbiome research, further automation, application of AI-driven data analysis, and continued improvements in dynamic range and throughput on next-generation mass spectrometers.

Conclusion

The BIOCRATES MxP Quant 500 kit on the Waters ACQUITY UPLC I-Class PLUS and Xevo TQ-XS platform offers a robust, high-throughput, and fully standardized targeted metabolomics solution with excellent accuracy, precision, and inter-laboratory comparability.

References

• U. Sommer, T. Koal, A. Peck, Standardized targeted metabolomics using the MxP Quant 500 kit on Waters ACQUITY UPLC I-Class PLUS and Xevo TQ-XS, Application Note, Waters Corporation, 2020.
• EMA and FDA bioanalytical method validation guidelines.
• NIST Standard Reference Material 1950 (Metabolites in Frozen Human Plasma).