Williams J P, Khan M U, Wong D
Department of Botany, University of Toronto, Ontario, Canada.
J Lipid Res. 1995 Jun;36(6):1407-12.
A simple technique is described for the analysis of positional distribution of fatty acids on di- and triacylglycerols using lipase and phospholipase A2 that de-esterify fatty acids from specific sn positions. The technique makes use of the fact that methanolic-NaOH methylates only fatty acids esterified to glycerol, while methanolic-HCl methylates both free and esterified fatty acids. After lipase action it is possible to determine the fatty acid released by lipase activity by comparing the fatty acid contents of the two methylation reactions. A computer program has been written to calculate enzyme activity and positional distribution from the results. The new technique is easier to use as it eliminates thin-layer chromatography used in the standard technique and can be performed on smaller samples using less lipase.
本文描述了一种简单的技术，用于分析甘油二酯和甘油三酯上脂肪酸的位置分布，该技术使用脂肪酶和磷脂酶A2从特定的sn位置去除脂肪酸酯。该技术利用了以下事实：甲醇-NaOH仅使酯化到甘油上的脂肪酸甲基化，而甲醇-HCl使游离脂肪酸和酯化脂肪酸都甲基化。脂肪酶作用后，可以通过比较两个甲基化反应的脂肪酸含量来确定脂肪酶活性释放的脂肪酸。已编写了一个计算机程序，根据结果计算酶活性和位置分布。这项新技术更易于使用，因为它省去了标准技术中使用的薄层色谱法，并且可以使用更少的脂肪酶对更小的样品进行操作。
