葡萄糖月桂酸脂的酶法合成与分离测定

Separation and analysis of lauroyl glucose ester synthesized by lipase

研究了葡萄糖月桂酸酯的酶法合成与分离测定。以丙酮为溶剂,用Novozym435脂肪酶催化合成了葡萄糖月桂酸酯,经液质联用(LC-MS)分析,反应产物为葡萄糖单酯和二酯,没有三酯或其它多酯生成。实验并优化了薄层色谱(TLC)法分析葡萄糖月桂酸酯的展开剂条件,得到展开剂∶甲苯∶乙酸乙酯∶甲醇∶水=10∶40∶4.5∶1.0(V/V/V/V),在此展开条件下,葡萄糖、葡萄糖单酯、二酯1和二酯2的Rf值相差均较大,分别为:0.098、0.51、0.85和0.97。用此展开剂作为流动相,采用硅胶柱层析分离葡萄糖月桂酸单酯,经高效液相色谱(HPLC)蒸发光散射检测(ELSD)和液质联用(LC-MS)鉴定,证实分离得到高纯度的葡萄糖月桂酸单酯。

The lauroyl glucose ester was synthesized in acetone with the Novozym 435 lipase as catalyst. The product was identified by HPLC-MS as gluose monoester and diesters,no triester or polyester was detected. The developing agent for thin layer chromatography （TLC） was tested and optimized. The final developing agent was the mixture of methylbenzene,acetic ester, methanol,and water with the volumic proportion of 10：40：4.5：1.0. With this developing agent,the R1 value of glucose, monester, diester 1 and diester 2 were respectively 0.098,0.51,0.85 and 0.9？. The developing agent was applied to silica gel column chromatography as mobile phase for separation and purification of lauroyl glucose esters. The purified monoester was tested by HPLC-ELSD and LC-MS, confirming that the purity of monoester 100%.