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本文介绍一种小型薄层层析系统,能迅速确定花生四烯酸盐代谢全部环氧化酶产物(即各种PGs)和几个最普通的脂氧化酶途径代谢产物(即各种羟基酸和白细胞三烯B_4),能分辨每点20μg的产物,相当于较大规模的薄层层析系统,一小时内能达到初步筛选全部主要的花生四烯酸盐代谢物。实验方法:含有被完整细胞和组织、匀浆,或亚细胞部分所转换的〔~(14)C〕花生四烯酸盐代谢产物的标本,用0.2M枸橼酸或0.5M醋酸酸化到pH3,用醋酸乙酯或乙醚提取两次。提取物用无水硫酸钠干燥,在氮气下脱水,以10μl乙醇或氯仿:甲醇(2∶1)溶液处理,加入前列腺素标准品(每种0.5～
This article presents a small, thin layer chromatography system that quickly identifies arachidonate metabolism of all cyclooxygenase products (ie, various PGs) and several of the most common lipid oxidase pathway metabolites (ie, various hydroxy acids And leukotriene B_4), capable of resolving 20 μg of product per point, is equivalent to a larger-scale TLC system that enables the primary screening of all major arachidonic acid metabolites within an hour. Experimental Methods: Specimens of [~ (14) C] arachidonate metabolites transformed with intact cells and tissues, homogenates, or subcellular fractions were acidified to pH 3 with 0.2 M citric acid or 0.5 M acetic acid , Extracted twice with ethyl acetate or ether. The extract was dried over anhydrous sodium sulfate, dehydrated under nitrogen, treated with 10 μl of ethanol or chloroform: methanol (2: 1) solution, prostaglandin standard (each 0.5 -