莱鲍迪苷A的制备色谱法分离纯化研究
Isolation and purification of Rebaoudioside-A by Medium Pressure Preparative Liquid Chromatography
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摘要: 本文从制备色谱填料入手,探讨采用中压制备色谱法一次分离得到高纯度莱鲍迪苷A(RA)的可行性。以羧酸改性15μm单分散PS-DVB基球填料为分离介质,装填入规格为12.6×2.66cm的flash色谱柱中,以RA单体的纯度、产率、回收率为考察指标,研究了不同的流动相比例、上样浓度、上样体积以及流动相流速对分离纯化过程的影响。建立了合适的操作条件:流动相为乙腈/水=80/20(v/v),上样浓度为280mg/mL,上样体积为0.4mL,流速为4mL/min。在最佳的操作条件下,分离得到的RA单体纯度为90.22%,回收率79%。该方法具有分离效率高,产品质量好,生产周期短,操作方便等特点,为从低含量甜菊糖结晶母液中纯化RA单体的工业化生产提供了一种有效的途径。Abstract: 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.