Isolation and purification of Rebaoudioside-A by Medium Pressure Preparative Liquid Chromatography

A medium pressure preparative liquid chromatography (MPLC) was used based on a type of packing material to isolate RA of high purity for the first time.The 15μm modified PS-DVB packing material was devised into a flash column (12.6 × 2.66cm) .Purity, yield rate and recovery rate were investigated to study the proportion of mobile phase, sample concentration, sample volume as well as the mobile phase flow rate on the separation and purification process of RA.The optimal operating conditions were as follows: the proportion of mobile phase (acetonitrile/water) was 80/20 (v/v) , sample concentration 280mg /mL, sample volume 0.4mL, and the flow rate 4mL /min.Under the optimal operating conditions, the obtained RA purity was 90.22%, and the recovery rate achieved 79%.This method was proved to be highly efficient for its good product quality, short production cycle, easy operation, providing a favorable reference for the RA industrialization research.