Garcés R, Mancha M
Instituto de la Grasa (CSIC), Sevilla, Spain.
Anal Biochem. 1993 May 15;211(1):139-43. doi: 10.1006/abio.1993.1244.
A procedure for the digestion of fresh tissue, transmethylation of lipids, and the extraction of fatty acid methyl esters (FAMES) in one step is described. Fresh tissues or isolated lipids are heated with a reagent containing methanol:heptane:benzene:2,2-dimethoxypropane:H2SO4 (37:36:20:5:2, by vol), methanol:heptane:toluene:2,2-dimethoxypropane:H2SO4 (39:34:20:5:2, by vol), or methanol:heptane:tetrahydrofuran:2,2-dimethoxypropane:H2SO4 (31:42:20:5:2; by vol). At 80 degrees C the simultaneous digestion and lipid transmethylation takes place in a single phase. After cooling at room temperature two phases are formed; the upper one contains the FAMES ready for GLC analysis. The procedure allows the determination of the fatty acid composition of lipids from tissues containing high proportions of triacylglycerols (oil seeds), water (leaves), or both (oil fruits). Due to the complete extraction of the lipids from the tissue, their content can be determined by adding an internal standard to the sample.
本文描述了一种一步消化新鲜组织、脂质转甲基化以及提取脂肪酸甲酯（FAMES）的方法。将新鲜组织或分离出的脂质与含有甲醇：庚烷：苯：2,2 - 二甲氧基丙烷：硫酸（体积比为37:36:20:5:2）、甲醇：庚烷：甲苯：2,2 - 二甲氧基丙烷：硫酸（体积比为39:34:20:5:2）或甲醇：庚烷：四氢呋喃：2,2 - 二甲氧基丙烷：硫酸（体积比为31:42:20:5:2）的试剂一起加热。在80℃时，消化和脂质转甲基化在单相中同时进行。室温冷却后形成两相；上层含有可供气相色谱分析的脂肪酸甲酯。该方法可用于测定含有高比例三酰甘油（油籽）、水（叶片）或两者皆有的组织（油果）中脂质的脂肪酸组成。由于能从组织中完全提取脂质，通过向样品中添加内标物可测定其含量。
