SPILL THE WINE: A RAPID MICROBORE METABOLIC PROFILING HILIC APPROACH COUPLED WITH ION MOBILITY HRMS FOR THE ANALYSIS OF ANTHOCYANINS IN RED WINE

Significance of the Topic

Anthocyanins are key flavonoid pigments responsible for the red, blue, and violet coloration of many plant-derived foods, notably red wine. Their chemical interactions with tannins influence wine aging, sensory properties, and stability. Rapid and detailed profiling of these compounds supports quality control, authenticity assessment, and research into wine chemistry.

Objectives and Article Overview

This study presents a rapid microbore HILIC method coupled with travelling wave ion mobility separation and high-resolution mass spectrometry (TW-IMS HRMS) to profile selected anthocyanins in retail red wines. The aim is to achieve efficient separation in under 4 minutes, resolve co-eluting species, and obtain accurate structural insight.

Methodology and Used Instrumentation

The analytical workflow combined microbore hydrophilic interaction chromatography with gas-phase ion mobility and high-resolution detection. Key UPLC parameters included:

• Column: BEH Amide, 1.7 µm, 1.0 x 50 mm, 50 °C
• Mobile phases: water + 0.1% formic acid (MP A); acetonitrile + 0.1% formic acid (MP B)
• Gradient elution and 0.2 µL injection volume; total run time 3.33 min

TW-IMS HRMS settings:

• Instrument: Waters Synapt G2-Si with low-flow ESI
• Polarity: positive ion mode, capillary 1.5 kV, cone 30 V
• Mass range: m/z 50–1200, scan time 0.1 s
• Ion mobility: wave height 40 V, velocity 650 m/s, nitrogen gas

Main Results and Discussion

Through replicated injections of diluted red wine samples, principal component analysis demonstrated reproducible clustering by wine variety. Eight anthocyanins were identified based on accurate mass measurements of precursors and fragments, retention times, and ion mobility separation. TW-IMS effectively resolved co-eluted compounds such as malvidin-3-O-glucoside pyruvic acid and malvidin-3,7-diglucoside in the gas phase, enhancing spectral clarity and confidence in structural assignment.

Benefits and Practical Applications

• High throughput profiling with sub-4-minute run times supports rapid screening.
• Ion mobility separation complements chromatography to differentiate isobaric or co-eluting species.
• Accurate mass measurement provides detailed structural information without extensive sample preparation.
• Applicability in quality control, authenticity verification, and metabolomic studies in enology.

Future Trends and Potential Applications

The integration of microbore HILIC-IMS HRMS can be extended to:

• Broader phenolic and metabolite profiling in other foods and beverages.
• Higher-throughput platforms and automated data analysis, including AI-driven interpretation.
• Targeted screening for vintage, varietal authentication, and geographical origin tracing.
• Combination with orthogonal separation techniques for comprehensive metabolomics.

Conclusion

This rapid microbore HILIC approach, combined with ion mobility and high-resolution MS, offers a robust, high-throughput solution for detailed anthocyanin profiling in red wine. The method enhances separation, structural insight, and reproducibility, benefiting both research and industrial applications.

Reference

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3. Gray G et al. (2016) Anal Chem 88:5742–5751
4. Calwineries. Chemical Components of Wine: Anthocyanins. Available online (accessed 2018).