Lipid Tandem Quadrupole Mass Spectrometer

脂质串联四极杆质谱仪

• 批准号: 7790416
• 负责人: ROBERT Carl MURPHY
• 金额: $ 35.89万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7790416
• 关键词: Anabolism; Apoptosis; Applications Grants; Arachidonic Acids; Arts; Back; Biochemical; Biochemical Pathway; Biochemistry; Cell Line; Cell physiology; Coculture Techniques; Colorado; Cytosolic Phospholipase A2; Disease; Fatty Acids; Goals; Health; High Pressure Liquid Chromatography; Infection; Investigation; Ions; Lead; Leukotrienes; Lipid Biochemistry; Lipids; Lung; Lung diseases; Lysophospholipids; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Metabolism; Methods; Mus; Mycoses; Pathway interactions; Phosphatidyl glycerol; Phosphatidylglycerols; Phosphatidylserines; Phospholipid Metabolism; Phospholipids; Physiological; Play; Process; Prostaglandins; Pulmonary Surfactants; Research; Research Personnel; Role; Running; Scanning; Transgenic Organisms; Universities; biological systems; follower of religion Jewish; instrument; instrumentation; lipid mediator; lysophosphatidylserine; macrophage; mass spectrometer; medical schools; multiple reaction monitoring; neoplastic cell; programs; public health relevance; quorum sensing; research study; response; stable isotope; tandem mass spectrometry

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.

描述（由申请人提供）：本资助申请的主要目的是为科罗拉多大学丹佛分校（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，以便通过准确、精确和敏感的定量分析促进脂质介质生物合成的研究；基础生物化学研究以磷脂代谢为中心。此外，参与细菌群体感应的脂质介质将在结构上进行阐明和定量。所要求的仪器是最先进的串联四极杆质谱仪，带有线性离子阱，可以进行多种质谱实验，包括前体离子扫描产物离子扫描，中性损失扫描，以及多种反应监测，MSn分析和直接质谱分析（MS分析）。质谱分析目前正在支持几个特定的研究项目，包括转基因和嵌合小鼠中白三烯的跨细胞生物合成的详细研究，巨噬细胞对真菌感染反应研究中产生的脂质介质的分析，以及胞质磷脂酶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，以及巨噬细胞和巨噬细胞系原代培养的各种肿瘤细胞共培养中花生四烯酸代谢物的定量研究。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景。这也可以被称为经济经济学。研究还建议支持磷脂生物化学的调查，包括在肺部感染期间磷脂酰甘油在肺表面活性剂中的作用，以及脂肪酸在被称为“Lands途径”的生化途径中再酰化回磷脂的作用。所要求的仪器具有大量多反应监测（MRM）实验的能力，不仅可以支持二十染色体组学研究，还可以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。该仪器将放置在我们的脂质质谱设备，并完全致力于脂质生化研究。