Simultaneous Determination of Hesperidin, Naringin and Their Aglycones in Processed Foods by Fast Semi-micro HPLC

A fast semi-micro liquid chromatographic method for the simultaneous determination of hesperidin, naringin and their aglycones, i. e, hesperetin and naringenin, in processed foods was developed. The sample was refiuxed with 60% aqueous methanol on a hot water bath at 90°C and cleaned up on a Sep-Pak C18 cartridge. The sample solution was chromatographed on a semi-micro column (2.1mm i. d×80mm) packed with 3μm particles of Capcell Pak C18-SG120 with a mobile phase of acetonitrile-water-0.2mol/L phosphate buffer, pH 4.0 (18:77:5). Detection was achieved with a UV monitor set at 283nm. The recoveries from soft drinks, marmalade, candy, yogurt, cookies and salad dressing fortified with the standard mixture at levels of 4 to 100μg/g were 82.6 to 100.4% (C. V. 1.6-5.2%) for hesperidin, 88.3 to 98.9% (C. V. 0.7-2.4%) for naringin, 88.3 to 93.4% (C. V. 1.3-3.9%) for hesperetin and 91.2 to 96.7% (C. V. 1.1-2.6%) for naringenin. The detection limit was 0.5μg/g for each compound.