Purification and Preparation of Rebaudioside A from Steviol Glycosides Using One-Dimensional Hydrophilic Interaction Chromatography

Abstract

A method for purifying and preparing rebaudioside A (RA) from steviol glycosides at preparative scale was developed with resin based on hydrophilic interaction liquid chromatography (HILIC). In pure water and acetone-water system, the adsorption capacity and selectivity of five anion resins for RA, rebaudioside C (RC) and stevioside (ST) were examined and discussed. Strongly basic IRA458 with the quaternary amine group was chosen as the resin used for separating and preparing RA. The hydroxide form of IRA458 (IRA458-OH) showed the best results in terms of the adsorption behaviors for RA, RC and ST. The retentions of RA, RC and ST on IRA458-OH resin at high concentration of acetone solution followed HILIC mode, in which retention is probably based on surface adsorption of the resin. Under optimized chromatographic conditions, the pooled purity and yield of RA were up to 98.18 and 98.62%, the relative standard deviations (n = 3) for these two parameters were 1.2 and 5.7%, respectively. The present method has the characteristics of simple, low cost, high purity and high yield. The study will be a promising tool for RA industrialized production and also provides a possible mode for purifying and preparing polar components from their analogs.