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.

Kinetics and thermodynamics of rebaudioside A adsorption on a strongly acidic cation exchange resin

“…The concentrations of samples before and after adsorption were detected with a Shimadzu LC-20AT HPLC instrument under the following chromatographic conditions [11]: column, Hypersil SAX (250 × 4.6 mm, 5 µm); mobile phase, acetonitrile-water (85:15, v/v); flow rate, 2.0 mL/min; column temperature, 30 • C; injection volume, 5 µL; and detector, UV (λ = 210 nm).…”

“…Empirically, quaternary ammonium groups are more hydrophilic than tertiary ammonium groups. As a consequence, SBA resins are significantly more hydrophilic than WBA resins [11,15]. Meanwhile, the matrix and pore structure also affect the hydrophilicity of the resin in the following order: styrenedivinylbenzene (DVB) < polystyrene < acrylic, macroporous < gel [11,16].…”

“…As a consequence, SBA resins are significantly more hydrophilic than WBA resins [11,15]. Meanwhile, the matrix and pore structure also affect the hydrophilicity of the resin in the following order: styrenedivinylbenzene (DVB) < polystyrene < acrylic, macroporous < gel [11,16]. In addition, compared with uncharged functional groups, the polar functional groups with positive or negative charges all display good hydrophilicity.…”

“…Currently, the main RA purification and preparation technologies reported in the literature include recrystallization, extraction, high-speed countercurrent chromatography, and resin-based methods [5][6][7][8][9][10][11]. Among them, the resin method has received increasing attention because of its inherent advantages.…”

“…Among them, the resin method has received increasing attention because of its inherent advantages. The reported studies focus on the separation and purification of RA using macroporous and anionic exchange resins [10,11]. Although the use of macroporous resins has allowed for high-purity RA preparation, they do not satisfy the requirements for industrial applications.…”