Simultaneous Quantification of Flavonoids and Biflavonoids in Ginkgo biloba Using RP-HPTLC Densitometry Method

A reversed-phase high-performance thin-layer chromatography (RP-HPTLC) method was developed for the determination of flavonoids (quercetin and kaempferol) and biflavonoids (sciadopitysin, ginkgetin, and bilobetin) in the aqueous methanolic extract of Ginkgo biloba. There have been a number of reports on the quantification of flavonoids in G. biloba using different analytical techniques, but the reported methods are not suitable for routine analysis and also for the simultaneous quantification of flavonoids and biflavonoids in G. biloba. The method employed here used precoated plates of silica gel 60F254 as the stationary phase with dual-run acetonitrile–water–methanol–formic acid (20:20:1:0.005, v/v/v/v) and acetonitrile–water–methanol–formic acid (20:17:1:0.005, v/v/v/v) as mobile phases with densitometric determination of flavonoids and biflavonoids at 254 nm in reflection/absorption mode. The linear regression analysis data for the calibration plots showed a linear relationship (r2 from 0.9706 to 0.9990). The method was validated for accuracy, precision, and robustness. The limits of detection (LOD) and quantification (LOQ) were in the range of 0.12–0.37 μg and 0.60–1.85 μg, respectively, for the analytes. The method is reproducible and convenient for quantitative analysis of these compounds in the leaves of G. biloba. This is the first report of simultaneous densitometry quantification of major flavonoids and biflavonoids in G. biloba using the newly developed and validated RP-HPTLC method