Quantitative estimation of hesperidin by HPTLC in different varieties of citrus peels

Objective: To develop a simple, selective, sensitive and accurate high-performance thin layer chromatography (HPTLC) method to determine the quantity of hesperidin in different varieties of citrus fruits. Methods: The method was carried out in aluminum-backed silica gel 60 F₂₅₄ plates with ethyl acetate-methanol-water 15:3:2 (%, v/v) as mobile phase. Results: A compact band was obtained for hesperidin at R_f value of (0.40±0.04). The calibration plot was linear in the range of 100-800 ng/spot of hesperidin and the correlation coefficient of 0.998 6 was indicative of good linear dependence of peak area on concentration. Limit of detection (8.87 ng/spot), limit of quantification (23.21 ng/spot), accuracy (less than 2%) and recovery (ranging from 98.55-99.38) were found satisfactory. Conclusions: The method developed can be used for routine analysis of hesperidin in crude drug as well as in herbal and pharmaceutical dosage form containing citrus fruits as an ingredient.