Rapid Simultaneous Analysis for Three Sugars Fraction in Enzymatic Degradation of Stachyose

The objective of the present study was to develop an effective method of High Performance Liquid Chromatography-Refractive Index Detector (HPLC-RID) for the simultaneous quantification of stachyose, raffinose and galactose in enzymatic degradation of stachyose. The three sugars were separated on an Agilent zorbax carbohydrate column with acetonitrile water (75:25 v/v). The calibration curves of the three standards exhibited good linearity (R 2 >0.9989). The Limits of Detection (LOD) values of the stachyose, raffinose and galactose were 0.137, 0.301 and 0.386 μg/mL, respectively and their Limits of Quantification (LOQ) values were 0.452, 0.993 and 1.25 μg/mL, respectively. The Relative Standard Deviations (RSDs) of the data of the intra- and inter-day experiments were less than 2.68, 2.42 and 3.70%, respectively. The results of the recovery test were 97.71-102.61% with an RSD value of 0.92-2.41%. In conclusion, the results of validation showed that the proposed HPLC-RID method was useful for the quantification of the three sugars in enzymatic degradation of stachyose and similar reaction.