Application of Evaporative Light Scattering Detector (Part 8) Analysis of Terpenoids in Ginkgo Biloba

Significance of the Topic

Ginkgo biloba terpenoids such as bilobalide and ginkgolides are bioactive compounds widely used in dietary supplements for improving cerebral and peripheral circulation. Their lack of chromophores makes evaporative light scattering detection combined with reversed-phase gradient HPLC an effective approach for reliable analysis.

Study Objectives and Overview

The primary aim was to develop and validate an HPLC method using an ELSD-LT II evaporative light scattering detector to separate and quantify four major terpenoids in Ginkgo biloba extract and commercial supplements.

Methodology and Instrumentation

An HPLC system equipped with a Shim-pack FC-ODS column (150 mm × 4.6 mm) operated at 50 °C was used. The mobile phase consisted of water (A) and methanol (B) with a gradient from 20 to 45 over 16 minutes, then to 80 between 16.01 and 20 minutes, returning to 20 by 30 minutes. The flow rate was 1.0 mL/min and injection volume 10 µL. Detection employed an ELSD-LT II with drift tube temperature at 40 °C, gain set to 6, and nitrogen nebulizer gas at 350 kPa.

Main Results and Discussion

A standard mixture of bilobalide and ginkgolides A, B, and C (200 mg/L each) yielded baseline separation in a single 30-minute run. Calibration over 20–1000 mg/L showed log-transformed linearity with correlation coefficients (R2) above 0.998 for all compounds. Repeatability at 200 mg/L achieved relative standard deviations below 1.6%.

Analysis of Commercial Supplement

Sample preparation involved tablet milling, methanol extraction with sonication, centrifugation, filtration, dilution, and 10 µL injection. The chromatogram of the dietary supplement confirmed the presence and quantitation of all four terpenoids with clear peak resolution.

Benefits and Practical Applications

• Allows sensitive detection of nonchromophoric terpenoids without derivatization
• Provides high reproducibility suitable for quality control in supplement manufacturing
• Employs straightforward sample preparation adaptable to routine laboratory workflows

Future Trends and Applications

Emerging directions include coupling ELSD with mass spectrometry for compound identification, development of enhanced aerosol detectors for increased sensitivity, miniaturized HPLC systems to reduce solvent use, and application of the technique to a wider range of nonchromophoric natural products.

Conclusion

The validated ELSD-HPLC method offers a robust, sensitive, and reproducible solution for quantifying Ginkgo biloba terpenoids in extracts and supplements, supporting stringent quality assurance and research needs.

References

Application News High Performance Liquid Chromatography No L391 Shimadzu Corporation Printed in Japan