A RAPID HPLC METHOD FOR THE DETERMINATION OF RAFFINOSE FAMILY OF OLIGOSACCHARIDES IN PEA-SEEDS

Two HPLC techniques for the determination of raffinose family oligosac charides (RFO - raffinose, stachyose and verbascose) in pea are descri bed. A reverse phase HPLC (RP-HPLC) on Silica C-18 column using demine ralized water as mobile phase enabled determination of the total conte nt of RFO within less than 10 minutes. Ion Moderated Partition HPLC (I MP-HPLC) using a strong cation exchanger, in calcium form, with demine ralized water as mobile phase allowed the determination of all the ind ividual saccharides within 25 minutes. In both RP-HPLC and IMP-HPLC re fractometric detection was used. The methods were compared on a repres entative pea sample series, and in both techniques the lowest detectab le concentration in sample solution was calculated to be 5 mg sacchari de/mL, i.e., the smallest detectable amount of each saccharide was 100 ng.