Screening Analysis of Metabolites in Red Wine

Significance of the Topic

The comprehensive profiling of metabolites in food matrices is essential for understanding nutritional value, taste characteristics and functional properties. High-resolution metabolomic screening helps research laboratories rapidly identify bioactive compounds, optimize quality control and support food science advancements.

Study Objectives and Overview

This application note illustrates the use of the Shimadzu LCMS-9030 quadrupole time-of-flight (Q-TOF) mass spectrometer coupled with UHPLC for rapid screening of metabolites in red wine. The primary goals were to demonstrate a streamlined workflow using LabSolutions Insight Explore™ software, prepare a candidate compound list from an endogenous metabolite database, and verify key metabolites such as gallic acid.

Methodology and Instrumentation

The workflow comprised the following steps:

• Sample preparation: Five red wine varieties were pooled, centrifuged and diluted tenfold with ultrapure water.
• Chromatographic separation: Nexera X3 UHPLC system with a reverse-phase column, 0.1% formic acid in water (A) and acetonitrile (B) gradient at 0.25 mL/min, column oven at 40 °C, 3 µL injection.
• Mass spectrometry: LCMS-9030 in negative ESI mode, data-dependent acquisition (DDA) to record precursor and MS/MS data. Key parameters included DL temperature 250 °C, interface 300 °C, CID gas 230 kPa.
• Data processing: A target list of ~500 primary metabolites (retention time, formula, exact mass) was loaded into LabSolutions Insight Explore. The Analyze function extracted peaks, and the Screen function matched peaks to candidate compounds within ±1 mDa mass error.

Main Results and Discussion

Analysis yielded a base peak chromatogram with 455 extracted peaks. Screening assigned 90 peaks to known metabolites, predominantly organic acids (citric, succinic, gallic acids) and flavonoids (quercetin 3-O-glucuronide). Gallic acid (m/z 169.0139) was identified with −0.35 mDa error and confirmed by matching retention time and MS/MS fragment patterns against a standard.

Benefits and Practical Applications

• The Q-TOF workflow enables high-throughput, accurate metabolite screening in complex food samples.
• LabSolutions Insight Explore simplifies target list creation and automated data analysis, reducing bottlenecks.
• Flexible targeting: any compound with known formula can be screened without custom method development.
• Applications include food quality control, nutraceutical research and functional food development.

Future Trends and Potential Applications

• Expansion of metabolite libraries with retention time and MS/MS databases to cover wider food and biological matrices.
• Integration of machine learning for automated annotation and predictive metabolite discovery.
• Coupling with other omics platforms (proteomics, lipidomics) for multi-layered food profiling.
• Application to real-time quality monitoring in production lines and on-site food authenticity testing.

Conclusion

The Shimadzu LCMS-9030 UPLC-Q-TOF system, combined with LabSolutions Insight Explore, provides an efficient and flexible platform for screening endogenous metabolites in red wine. The approach successfully identified key organic acids and flavonoids, with gallic acid confirmation demonstrating quantitative accuracy. This rapid workflow supports diverse food analysis and metabolomics research needs.

Reference

Y. Umakoshi and T. Iida. Screening Analysis of Metabolites in Red Wine. Shimadzu Application News, First Edition: Mar. 2022