Lipid Tandem Quadrupole Mass Spectrometer

描述（由申请人提供）：本资助申请的主要目的是为科罗拉多丹佛大学（医学院）和国家犹太健康中心的研究人员提供最先进的电喷雾串联质谱，以促进通过准确、精确和灵敏的定量分析研究脂质介质生物合成;以及围绕磷脂代谢的基础生化研究。此外，脂质介质，参与细菌群体感应将在结构上阐明和定量。所要求的仪器是一个国家的最先进的串联四极杆质谱仪与线性离子阱，允许几个质谱实验进行，包括前体离子扫描产品离子扫描，中性损失扫描，以及多反应监测，MSn分析，和直接质谱分析（MS分析）。几个具体的研究计划目前正在支持质谱分析，包括在转基因和嵌合小鼠的白三烯的跨细胞生物合成的详细研究，在巨噬细胞对真菌感染的反应的研究中产生的脂质介质的分析，和胞质磷脂酶A2的作用。与细胞凋亡相关的生物活性磷脂酰丝氨酸和溶血磷脂酰丝氨酸代谢物的结构表征，以及在各种肿瘤细胞与巨噬细胞和巨噬细胞系原代培养物共培养研究中产生的花生四烯酸代谢物的定量。这些研究的主要目标是了解脂质介质在健康和疾病（包括癌症）中的作用，并涉及在单次HPLC运行中定量测量20 - 30种不同的花生四烯酸代谢物，以获得特定生物系统中花生四烯酸代谢的完整图像。这也可以被称为eicosanomics。还提出了支持磷脂生物化学研究的研究，包括肺感染期间磷脂酰甘油在肺表面活性物质中的作用，以及在称为"Lands途径"的生物化学途径中脂肪酸再酰化回磷脂。“所要求的仪器具有大量多反应监测（MRM）实验的能力，这些实验不仅将支持二十碳纳米管组学研究，还将支持与关键细胞过程以及肺部疾病相关的详细磷脂和溶血磷脂生化研究。该仪器将被放置在我们的脂质质谱设施，并完全致力于脂质生化研究。 公共卫生相关性：已知花生四烯酸的代谢产物通过形成胡萝卜素和白三烯在健康和疾病中起重要作用。为了定量测量生物系统中的这些分子，使用稳定同位素稀释和串联质谱的方法是首选的方法。除此之外，详细的生化研究， 磷脂和溶血磷脂的研究将揭示以前没有考虑过的脂质生物化学的基本生物化学机制。这些研究将使我们对正常生理和病理生理过程的理解有所突破。

DESCRIPTION (provided by applicant): The broad objective of this grant application is to make available state-of-the-art electrospray tandem mass spectrometry to investigators at the University of Colorado Denver (School of Medicine) and National Jewish Health in order to facilitate studies of lipid mediator biosynthesis through accurate, precise, and sensitive quantitative analysis; and basic biochemical studies centered around phospholipid metabolism. In addition, lipid mediators that are involved in bacterial quorum sensing will be structurally elucidated and quantitated. The instrumentation requested is a state-of-the-art tandem quadrupole mass spectrometer with a linear ion trap that permits several mass spectrometric experiments to be carried out, including precursor ion scanning product ion scanning, neutral loss scanning, as well as multiple reaction monitoring, MSn analysis, and direct mass spectrometric analysis (MS analysis). Several specific research programs are currently being supported by mass spectrometric analysis including the detailed studies of transcellular biosynthesis of leukotrienes in transgenic and chimeric mice, analysis of lipid mediators generated in studies of macrophage response to fungal infections, and the role of cytosolic phospholipase A2. Structural characterization of biologically active phosphatidylserine and lysophosphatidylserine metabolites relevant to cellular apoptosis, and quantitation of metabolites of arachidonic acid generated in studies of coculture of various tumor cells with primary cultures of macrophages and macrophage cell lines. These studies have the primary goal of understanding the role of lipid mediators in health and disease, including cancer, and involve the quantitative measurement of 20-30 different metabolites of arachidonic acid in a single HPLC run in order to obtain a complete picture of the metabolism of arachidonic acid within a specific biological system. This could also be called eicosanomics. Studies are also proposed to support investigation of phospholipid biochemistry, including the role of phosphatidylglycerol in pulmonary surfactant during pulmonary infections as well as reacylation of fatty acids back into phospholipids in a biochemical pathway termed the "Lands Pathway." The instrument requested has the capability of a large number of multiple reaction monitoring (MRM) experiments that will support not only the eicosanomics studies, but also the detailed phospholipid and lysophospholipid biochemical studies related to critical cellular processes as well as pulmonary disease. The instrument will be placed in our lipid mass spectrometry facility and devoted entirely to lipid biochemical investigations. PUBLIC HEALTH RELEVANCE: Metabolites of arachidonic acid are known to play important roles in health and disease through the formation of prostaglandins and leukotrienes. In order to quantitatively measure these molecules in biological systems, the approach using stable isotope dilution and tandem mass spectrometry is the method of choice. In addition to this, detailed biochemical studies of phospholipid and lysophospholipids can be pursued that will reveal basic biochemical mechanisms of lipid biochemistry not previously considered. These studies will lead to breakthroughs in our understanding of normal physiological as well as pathophysiologic processes.