Densitometric-High-Performance Thin-Layer Chromatographic Estimation of Diosmin, Hesperidin, and Ascorbic Acid in Pharmaceutical Formulations

An attempt was made to develop and validate a simple, accurate high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of diosmin, hesperidin, and ascorbic acid in pharmaceutical preparations. Analysis was performed on 10 cm × 20 cm aluminum-backed plates coated with 0.2 mm layers of silica gel 60 F254 (Merck, Germany). CAMAG TLC Scanner III was used for UV densitometric scanning. The used HPTLC system was found to give sharp, symmetrical, and well-resolved peaks at RF values of 0.34 ± 0.02, 0.40 ± 0.04, and 0.56 ± 0.03, and linearity was found in the ranges 100–800 ng/spot (r2 = 0.9985), 100–800 ng/spot (r2 = 0.9966), and 50–400 ng/spot (r2 = 0.9951) for diosmin, hesperidin, and ascorbic acid, respectively.