Fatty Acid and Eicosanoid Analysis Core/Robert C. Murphy

脂肪酸和类二十烷酸分析核心/Robert C. Murphy

• 批准号: 8007062
• 负责人: FLOYD H CHILTON
• 金额: $ 13.45万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8007062
• 关键词: Arachidonate 15-Lipoxygenase; Arachidonic Acids; Arts; Biochemical; Biological; Biological Assay; Biological Models; Blood capillaries; Botanicals; Cells; Coupled; Development; Eicosanoids; Eicosapentaenoic Acid; Event; Fatty Acids; Health; Individual; Investigation; Knowledge; Lead; Leukotrienes; Lipids; Liquid Chromatography; Mass Fragmentography; Mass Spectrum Analysis; Measures; Oils; Pathway interactions; Polyunsaturated Fatty Acids; Preventive; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Research Project Grants; Sampling; Sensitivity and Specificity; Techniques; Technology; base; capillary; ionization; liquid chromatography mass spectrometry; research study; tandem mass spectrometry; tool

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.

很明显，为了了解发生的生化事件的复杂性 在导致前列腺素和白三烯等类二十烷酸合成的细胞内，必须具有 一种可以测量这些途径的几乎所有产物的方法。分析学的发展 技术与我们对这些需求的理解保持同步，因此非常强大的工具 现在可用。质谱分析法已成为基于灵敏度和稳定性的首选技术。 本申请中提出的项目所要求的具体性。电喷雾电离和电喷雾技术的进展 串联质谱法与液相色谱法 (LC/MS) 结合使用时，可以进行 在一次 LC/MS/MS 实验中提取 30 种或更多目标类二十烷酸。毛细管气体 色谱/质谱 (GC/MS) 仍然是一种无与伦比的技术，能够识别 并定量脂肪酸（一次实验中多达 20 种脂肪酸）。基于这一技术水平 技术，该核心的主要具体目标是提供最先进的目标脂质检测方法 例行基础，以促进中心提案内每个单独研究项目的调查。 具体来说：1）液相色谱/质谱（LC/MS）和串联质谱 (LC/MS/MS) 将用于定量花生四烯酸和二十碳五烯酸的代谢物 环加氧酶和 5- 和 12/15-脂加氧酶途径以及 2) 气相色谱/质谱法 (GC/MS) 将用于定量和鉴定脂肪酸，包括生物制品中的多不饱和脂肪酸 提取物。