High-Speed Analysis of Glycyrrhizinate

Importance of the Topic

Glycyrrhizin is a principal bioactive triterpene glycoside found in licorice (Glycyrrhiza glabra) with significant applications in food, pharmaceuticals, and cosmetics. Rapid and reliable quantification is essential for quality control, safety assessment, and standardization of licorice-derived products.

Objectives and Study Overview

This study aimed to develop a high-speed liquid chromatography–mass spectrometry (LC-MS) method for sensitive and efficient determination of glycyrrhizin in licorice samples. Emphasis was placed on reducing analysis time while maintaining resolution and reproducibility.

Methodology and Instrumentation

• Sample Preparation: 100 mg licorice powder extracted with 10 mL water at 60 °C for 20 minutes, centrifuged and filtered through a 0.45 µm membrane.
• Chromatographic Conditions: Shim-pack XR-ODS column (75 mm × 2.0 mm i.d.), gradient elution from 25 % to 35 % acetonitrile in 50 mmol/L ammonium acetate (pH 4.7) over 4 minutes, flow rate 0.6 mL/min, column temperature 40 °C.
• MS Detection: Electrospray ionization in negative mode (ESI-), full-scan monitoring of glycyrrhizin ion at m/z 821.
• Injection Volume: 2 µL of filtered extract.

Used Instrumentation

• Prominence UFLC system coupled with LCMS-2010EV (Shimadzu).
• Shim-pack XR-ODS analytical column.
• Standard laboratory equipment for extraction, centrifugation, and filtration.

Main Results and Discussion

The method enabled clear separation of glycyrrhizin with a retention time of under 5 minutes. The extracted ion chromatogram at m/z 821 demonstrated a single, well-defined peak corresponding to glycyrrhizin. The rapid gradient and optimized conditions achieved high throughput without sacrificing sensitivity or peak shape. Detailed signal-to-noise evaluations indicated limits of detection in the low microgram per milliliter range.

Benefits and Practical Applications

This high-speed LC-MS protocol offers a streamlined workflow for routine analysis in quality assurance and regulatory laboratories. Its rapid turnaround supports large batch testing in the nutraceutical, pharmaceutical, and food industries. The negative ESI approach ensures selectivity for acidic glycosides amid complex plant matrices.

Future Trends and Potential Uses

Further integration with automated sample handling and miniaturized columns could reduce solvent consumption and increase throughput. Coupling with high-resolution mass spectrometry or tandem MS may broaden identification of minor licorice constituents. Method adaptation for other triterpene glycosides can expand applications across botanical analysis.

Conclusion

The presented LC-MS method provides a fast, sensitive, and reproducible approach for quantifying glycyrrhizin in licorice. Its combination of short run time and robust detection makes it highly suitable for quality control environments and research laboratories.

References

• Shimadzu Application Data Sheet LAAN-A-LC-E097, High-Speed Analysis of Glycyrrhizinate, October 2007