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 d etermination of hesperidin, naringin and their aglycones, i.e, hespere tin and naringenin, in processed foods was developed. The sample was r efluxed with 60% aqueous methanol on a hot water bath at 90 degrees C and cleaned up on a Sep-Pak C-18 cartridge. The sample solution was ch romatographed on a semi-micro column (2.1 mm i.d x 80 mm) packed with 3 mu m particles of Capcell Pak C-18-SG120 with a mobile phase of acet onitrile-water-0.2 mol/L phosphate buffer, pH 4.0 (18:77:5). Detection was achieved with a UV monitor set at 283 nm. The recoveries from sof t drinks, marmalade, candy, yogurt, cookies and salad dressing fortifi ed with the standard mixture at levels of 4 to 100 mu g/g were 82.6 to 100.4% (C.V. 1.6 similar to 5.2%) for hesperidin, 88.3 to 98.9% (C.V. 0.7 similar to 2.4%) for naringin, 88.3 to 93.4% (C.V. 1.3 similar to 3.9%) for hesperetin and 91.2 to 96.7% (C.V. 1.1 similar to 2.6%) for naringenin. The detection limit was 0.5 mu g/g for each compound.