High-performance thin-layer chromatography method for the simultaneous determination of vulgarin and epivulgarin biomarkers in artemisia judaica L

Novel, simple, and sensitive high-performance thin-layer chromatography (HPTLC) with fluorescence detection method has been successfully established and validated for the simultaneous determination of vulgarin and epivulgarin in different collections of Artemisia judaica. HPTLC method was carried out using glass plates coated with silica gel 60 F254 using petroleum ether-acetone (7:3, v/v) as the mobile phase. After development, the plates were scanned and quantified densitometrically at 224 nm for both vulgarin and epivulgarin. The two compounds' peaks from A. judaica were identified by comparing their single spot at RF = 0.30 ± 0.02 and RF = 0.36 ± 0.01, respectively, with those of vulgarin and epivulgarin. Linear regression analysis revealed a good linear relationship between peak area and amount of vulgarin and epivulgarin in the range of 100-700 ng band-1 for both compounds. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines, for precision, accuracy, and robustness. The proposed method will be useful to measure the therapeutic dose of vulgarin and epivulgarin in A. judaica extracts.