Simultaneous Quantification of Six Polymethoxyflavones in Gardenia lucida Roxb. Using High-Performance Thin-Layer Chromatography

摘要

Dikamali (Gardenia lucida Roxb., Rubiaceae family) is found to possess various ethno-medical properties, viz., anti-inflammatory, anthelmintic, cleaning foul ulcers, and wounds, to keep away flies from wounds in animals. In spite of having various medicinal properties, no validated analytical method has been reported so far for the quality assessment of Dikamali. Thus, a high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of six polymethoxyfl avones (PMFs) isolated from G. lucida. Chemical markers, e.g., gardenin-E (1), gardenin-D (2), xanthomicrol (3), 5-desmethynobiletin (4), gardenin-A (5), and gardenin-B (6), of G. lucida gum were selected for quality assessment. For the quantitative determination of the marker compounds, silica gel 60 F-254 TLC plates and solvent system n-hexane. diethyl ether. 1,4-dioxane (4:6:1, v/v) were used for optimum separation and selective evaluation. Densitometric determination was done at 335 nm for the targeted PMFs (1-6). Optimized chromatographic conditions provide well-separated compact bands for the tested polymethoxyfl avones. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.06 to 0.10 mu g and 0.21 to 0.32 mu g, respectively. Curves were linear from 0.33 to 1.67 mu g spot(-1) with acceptable correlation coefficients ranging from 0.9972 to 0.9988 of all PMFs. The percent mean recoveries of the PMFs ranged from 96.01 to 99.30. The analysis of three G. lucida samples from different locations verified the method to be reproducible and convenient. The method was validated as stated by the International Conference on Harmonization (ICH) guidelines. The developed HPTLC method is found to be accurate and precise. The same has successfully been applied for the estimation of the targeted PMFs in G. lucida gum samples of different locations.