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 Rf 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.