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 R_F 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 (r ² = 0.9985), 100-800 ng/spot (r² = 0.9966), and 50-400 ng/spot (r² = 0.9951) for diosmin, hesperidin, and ascorbic acid, respectively.