桔皮中橙皮苷的新型双水相萃取

Hesperidin Extraction from Orange Peel with New Aqueous Two-Phase System

采用新型双水相体系对桔皮中橙皮苷进行分离、纯化研究,考察了粗提物用量、pH、温度等参数对橙皮苷在两相间的分配及其回收率的影响。首先选取2种低分子有机物及3种无机盐分别构成双水相体系,用浊点法对其成相性能进行研究。选取其中成相性能较好的物质构成双水相体系,对橙皮苷的乙醇粗提物进行双水相分离,考察体系及其成相浓度对橙皮苷分配系数的影响。用所得最佳体系——丙酮-K2HPO4双水相体系对桔皮中橙皮苷进行分离。最佳条件下,橙皮苷的萃取收率可达96.69%。采用制备型HPLC对其进一步纯化,梯度洗脱时分离效果较好。经测定,双水相体系纯化后,橙皮苷样品纯度约为71.36%。经制备型HPLC精制所得橙皮苷纯度为96.09%。橙皮苷总回收率为83.54%。

In this manuscript,the separation and purification process of hesperidin from orange peel were developed,through investigating the effect of crude extract volume,pH and temperature on the variation of hesperidin partition and its recovery. Firstly two organic compounds with low molecular weight and three inorganic salts were selected to construct the aqueous two-phase system,whose performance was characterized by cloud point method. The chosen systems with good property in phase forming were applied to extract hesperidin from its crude ethanol extract, meanwhile the dependence of partition coefficient on the concentration of phase-forming substance and the type of system were also researched here. Therefore the best two-phase system was obtained,which is Acetone-K2HPO4 and applied to extract hesperidin from the peels. The recovery of hesperidin could reach to 96.69% under the optimization condition. The extract was refined further with prep-HPLC and purer hesperidin product was received. Gradient elution strategy made the refining process more effective in prep-HPLC system. The purity of hesperidin samples obtained from aqueous two-phase extraction was about 71.36%. It increased to 96.09% ca. after pre-HPLC refining. The total recovery of hesperidin was 83.54%.