High-speed analysis of Xanthohumol

Importance of the Topic

Rapid and accurate quantification of hop-derived flavonoids such as xanthohumol is essential for quality control in the brewing industry. Xanthohumol possesses antioxidant and health-promoting properties, making its monitoring valuable for product standardization and regulatory compliance.

Study Objectives and Overview

This application note describes a high-speed reversed-phase LC method for simultaneous determination of key hop compounds—humulinones, iso-α-acids, isoxanthohumol, α-acids, β-acids, and xanthohumol—in beer and raw hop extracts. The goal is to achieve efficient separation, reliable quantification, and suitability for routine brewery analysis.

Methodology and Instrumentation

Column: Shim-pack Velox C18 (50 mm × 3.0 mm, 1.8 μm)
System: Nexera X3 UHPLC with PDA detector (SPD-M40)
Mobile Phase A: 10 mmol/L sodium phosphate buffer (pH 2.6) with 0.2 mmol/L EDTA
Mobile Phase B: Methanol
Gradient: 50% B at 0 min → 90% B at 6 min → hold to 7 min → revert to 50% at 7.01 min; total run time 8 min
Flow Rate: 0.7 mL/min; Column Temp: 40 °C; Injection Volume: 5 μL

Main Results and Discussion

The method achieved baseline separation of six analytes within eight minutes. Retention times were reproducible (<0.5% RSD), and calibration curves exhibited excellent linearity (R² > 0.999). Limits of detection ranged from 0.02 to 0.05 mg/L, sufficient for trace-level monitoring in beer matrices. Precision and accuracy tests showed recoveries between 95% and 105% across concentrations relevant to European Brewery Convention guidelines.

Benefits and Practical Applications

• High throughput analysis: under 8 min per run enables large sample batches.
• Robust separation: clear resolution of structurally similar hop compounds.
• Trace-level sensitivity: meets brewery QA/QC and research needs.
• Broad applicability: suitable for both finished beer and hop extract testing.

Future Trends and Applications

Advances in column technology and detector sensitivity can further reduce run times and detection limits. Integration with automated sample prep systems will enhance throughput. The method may be extended to profiling additional bioactive prenylated flavonoids and tracking their transformations during brewing or storage.

Conclusion

This high-speed UHPLC method using Shim-pack Velox C18 and Nexera X3 offers a streamlined solution for simultaneous quantification of xanthohumol and related hop constituents. Its speed, sensitivity, and reproducibility make it a valuable tool for brewery quality control and research on health-related hop components.