Lehmann W D, Metzger K, Stephan M, Wittig U, Zalán I, Habenicht A J, Fürstenberger G
German Cancer Research Center, Heidelberg.
Anal Biochem. 1995 Jan 1;224(1):227-34. doi: 10.1006/abio.1995.1034.
An assay for the quantitative determination of the hydroxylation profile of long-chain fatty acids is described for gas chromatography negative-ion chemical ionization mass spectrometry and stable isotope dilution using [carboxyl-18O2]-labeled internal standards. The assay has been applied to the study of fatty acids isolated from body fluids, tissue, and cultured cells. Examples for the analyses of biological systems expressing 5-, 8-, 12-, or 15-lipoxygenase activity are given and the most important sources of analytical errors are addressed. Increased specificity compared to analysis by negative-ion chemical ionization, at the cost of sensitivity, can be achieved by the use of positive-ion electron impact ionization for the investigation of hydrogenated pentafluorobenzylester/trimethylsilylether derivatives. The method described provides complete, specific, and quantitative profiles of hydroxylated fatty acids originally present in biological samples or generated in vitro by incubation with polyunsaturated fatty acid substrates such as linoleic or arachidonic acid.
本文描述了一种用于定量测定长链脂肪酸羟基化谱的分析方法，该方法采用气相色谱负离子化学电离质谱法，并使用[羧基 - 18O2]标记的内标进行稳定同位素稀释。该分析方法已应用于从体液、组织和培养细胞中分离出的脂肪酸的研究。文中给出了表达5 -、8 -、12 - 或15 - 脂氧合酶活性的生物系统的分析示例，并讨论了最重要的分析误差来源。与负离子化学电离分析相比，通过使用正离子电子轰击电离来研究氢化五氟苄酯/三甲基硅醚衍生物，可在牺牲灵敏度的情况下提高特异性。所述方法可提供生物样品中原本存在的或通过与多不饱和脂肪酸底物（如亚油酸或花生四烯酸）孵育在体外生成的羟基化脂肪酸的完整、特异和定量谱图。
