A Rapid HPLC Method for the Determination of Raffinose Family of Oligosaccharides in Pea Seeds

SUMMARY Two HPLC techniques for the determination of raffinose family oligosaccharides (RFO - raffinose, stachyose and verbascose) in pea are described. A reverse phase HPLC (RP-HPLC) on Silica C18 column using demineralized water as mobile phase enabled determination of the total content of RFO within less than 10 minutes. Ion Moderated Partition HPLC (IMP-HPLC) using a strong cation exchanger, in calcium form, with demineralized water as mobile phase allowed the determination of all the individual saccharides within 25 minutes. In both RP-HPLC and IMP-HPLC refractometric detection was used. The methods were compared on a representative pea sample series, and in both techniques the lowest detectable concentration in sample solution was calculated to be 5 mg saccharide/mL, i.e., the smallest detectable amount of each saccharide was 100 ng.