Fatty Acid and Eicosanoid Analysis Core/Robert C. Murphy

很明显，为了理解发生的生化事件的复杂性 在导致前列腺素和白三烯等二十烷类化合物合成的细胞内，必须有 一种可以测量这些途径的几乎所有产物的方法。分析方法的发展 技术已经跟上了我们对这些需求的理解，因此非常强大的工具 现已上市。质谱学已经成为基于灵敏度和 本申请中提出的项目所要求的专用性。电喷雾电离技术的研究进展 串联质谱仪与液相色谱(LC/MS)联用，使携带 在一次LC/MS/MS实验中，检出了30个目标二十烷类化合物或更多。毛细管气 气相色谱/质谱仪(GC/MS)仍然是一种无与伦比的鉴定技术。 并对脂肪酸进行定量(一次实验中多达20种脂肪酸)。基于这种最先进的技术 技术，这个核心的主要具体目标是提供最先进的脂类检测方法。 在常规基础上，为中心提案内的每个单独研究项目的调查提供便利。 具体：1)液-质联用(LC/MS)和串联质谱仪 (LC/MS/MS)将用于定量花生四烯酸和二十碳五烯酸的代谢物。 环氧合酶和5-和12/15-脂氧合酶途径及2)气相色谱/质谱仪 (GC/MS)将用于定量和鉴定脂肪酸，包括生物中的多不饱和脂肪酸 萃取物。

It has become quite clear that in order to understand the intricacies of biochemical events taking place within cells that lead to the synthesis of eicosanoids such as prostaglandins and leukotrienes, one must have a means by which one can measure virtually all products of these pathways. The development of analytical techniques have kept pace with our understanding of these needs so that extraordinarily powerful tools are now available. Mass spectrometry has emerged as the technique of choice based on sensitivity and specificity demanded by the projects proposed in this application. Advances in electro-spray ionization and tandem mass spectrometry when coupled to liquid chromatography (LC/MS), has made it possible to carry out 30 targeted eicosanoids, or more, in a single LC/MS/MS experiment. Capillary gas chromatography/mass spectrometry (GC/MS) remains an unparalleled technique in its capability to identify and quantitate fatty acids (as many a 20 fatty acids in a single experiment). Based on this state of the art technology, the major specific aims of this core are to provide state of the art assays for target lipids on a routine basis to facilitate investigations in each of the individual research projects within the Center proposal. Specifically: 1) liquid chromatography/mass spectrometry (LC/MS) and tandem mass spectrometry (LC/MS/MS) will be used to quantitate metabolites of arachidonic acid and eicosapentaenoic acid by the cyclooxygenase and 5- and 12/15-lipoxygenase pathway and 2) Gas chromatography/mass spectrometry (GC/MS) will be used to quantitate and identify fatty acids, including polyunsaturated fatty acids in biological extracts.