Yamane M, Abe A, Yamane S
Department of Biochemistry, Tokyo Medical College, Japan.
J Chromatogr. 1994 Feb 11;652(2):123-36. doi: 10.1016/0378-4347(93)e0394-6.
A method for the analysis of epoxy polyunsaturated fatty acids (EpPUFAs) and epoxyhydroxy polyunsaturated fatty acids (EpHPUFAs) in rat tissue homogenate, with homo-gamma-linolenic acid (20:3, n - 6), arachidonic acid (20:4, n - 6), eicosapentaenoic acid (20:5, n - 3) or docosahexaenoic acid (22:6, n - 3) as a substrate, has been developed. Extraction with dichloromethane at pH 4-5 and concentration in the presence of pyridine were performed. Spectral analysis of chromatograms obtained with high-performance liquid chromatography-thermospray mass spectrometry showed the presence of EpPUFAs, EpHPUFAs and dihydroxy metabolites (DiHPUFAs) of EpPUFAs corresponding to each precursor fatty acid. On a selected-ion monitoring chromatogram, many EpPUFAs, EpHPUFAs and DiHPUFAs in an extract from an incubation mixture of each precursor fatty acid in aged rat tissue homogenate were detected simultaneously within 70 min. EpPUFAs and DiHPUFAs derived from 20:3 (n - 6) or 20:5 (n - 3) were detected in significant amounts. From these results, a highly active cytochrome P450 system or non-enzymic oxidative reactions in aged rat tissue homogenate were suggested.
已开发出一种分析大鼠组织匀浆中环氧多不饱和脂肪酸（EpPUFAs）和环氧羟基多不饱和脂肪酸（EpHPUFAs）的方法，该方法以同型γ-亚麻酸（20:3，n - 6）、花生四烯酸（20:4，n - 6）、二十碳五烯酸（20:5，n - 3）或二十二碳六烯酸（22:6，n - 3）作为底物。采用在pH 4 - 5条件下用二氯甲烷萃取，并在吡啶存在下进行浓缩。用高效液相色谱 - 热喷雾质谱法获得的色谱图的光谱分析表明，存在与每种前体脂肪酸相对应的EpPUFAs、EpHPUFAs和EpPUFAs的二羟基代谢物（DiHPUFAs）。在选择离子监测色谱图上，在70分钟内可同时检测到老年大鼠组织匀浆中每种前体脂肪酸孵育混合物提取物中的许多EpPUFAs、EpHPUFAs和DiHPUFAs。检测到大量源自20:3（n - 6）或20:5（n - 3）的EpPUFAs和DiHPUFAs。从这些结果推测，老年大鼠组织匀浆中存在高活性细胞色素P450系统或非酶促氧化反应。
