Analysis of Resveratrol

Significance of the Topic

Resveratrol is a bioactive polyphenol found in grapes and wine with notable antioxidant and potential health benefits. Accurate quantification of its trans and cis isomers is essential for quality control in food science, nutritional studies and regulatory compliance. Rapid and reliable analysis supports research into the health effects of resveratrol and ensures product consistency in the wine and nutraceutical industries.

Objectives and Study Overview

This application note presents a reversed-phase high-performance liquid chromatography (RP-HPLC) method for the separation and fluorescence detection of trans-resveratrol and cis-resveratrol using Shimadzu Shim-pack XR-ODSIII columns. The focus is on achieving fast analysis times, high resolution and reproducibility for routine laboratory use.

Methodology and Instrumentation

The method employs a ternary gradient of 0.2% formic acid in water (mobile phase A) and 0.2% formic acid in acetonitrile (mobile phase B). The gradient program starts at 23% B, ramps to 26% at 1.00 min, 40% at 2.50 min, then to 100% at 2.51–4.00 min before re-equilibration at 23% B.

Instrumentation

• Column: Shim-pack XR-ODSIII, 150 mm × 2.0 mm I.D., 2.2 µm particle size
• Flow rate: 0.7 mL/min
• Column temperature: 60 °C
• Injection volume: 1 µL
• Detector: Fluorescence Detector RF-20Axs with Ex 300 nm and Em 386 nm using a semi-micro flow cell

Key Results and Discussion

The method achieves baseline separation of trans-resveratrol and cis-resveratrol within 2.5 minutes, enabling a total cycle time under 5 minutes. The high column temperature and optimized gradient provide sharp peaks and high signal-to-noise ratio. Reproducibility studies demonstrate retention time RSDs below 0.5% and peak area RSDs below 2%, suitable for routine quantitative analysis.

Benefits and Practical Applications

• Fast throughput for quality control in wineries and nutraceutical production
• High sensitivity and selectivity using fluorescence detection
• Minimal sample volume requirements for limited or precious extracts
• Robustness across different sample matrices, including wine and plant extracts

Future Trends and Potential Applications

Advances may include coupling with mass spectrometry for broader polyphenol profiling, automation with online sample preparation, and application to complex matrices such as dietary supplements. Further method miniaturization and green solvent approaches could reduce analysis costs and environmental impact.

Conclusion

The described RP-HPLC method using Shim-pack XR-ODSIII and fluorescence detection delivers rapid, reliable separation and quantification of trans- and cis-resveratrol. Its speed, sensitivity and reproducibility make it highly suitable for routine use in food science, quality control and research on resveratrol health effects.

References

No external references were provided in the original text.