Rapid Validated RP-HPTLC Method for the Quantification of Major Bioactive Constituents of Crataegus oxyacantha L.

A reverse phase high-performance thin-layer chromatography (RPHPTLC) method was developed for determination of vitexin, hyperoside, vitexin-2"-O-rhamnoside, quercetin, and apigenin in the Crataegus oxyacantha extract. The method employed precoated plate of RP-18 silica gel 60F₂₅₄ as the stationary phase with acetonitrile-methanol-water-formic acid (10:10:20:0.05, v/v) as mobile phase, and densitometric determination was carried out at wavelength 254 nm in reflection/absorption mode. The linear regression analysis data for the calibration plots showed linear relationship (r) from 0.9985 to 0.9992. The method was validated for accuracy, precision, and robustness. The limits of detection and quantification were in the range of 100-300 ng and 310-960 ng, respectively, for analytes. The method is reproducible and convenient for quantitative analysis of these flavonoids in the leaves of C. oxyacantha. This is the first report of simultaneous densitometry quantification of major bioactive constituents in C. oxyacantha by developed and validated RP-HPTLC method. The change in the content of bioactive constituents with growth of the plant was also examined in leaf samples collected for 2 years. [ABSTRACT FROM AUTHOR]