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）：本赠款申请的广泛目标是为科罗拉多大学丹佛大学（医学院）和国家犹太人健康的研究人员提供最先进的电喷雾串联质谱，以促进通过准确的，精确的，敏感的定量分析来研究脂质介体生物合成的研究；以及基本的生化研究以磷脂代谢为中心。此外，与细菌群体相关的脂质介质将在结构上阐明和定量。所请求的仪器是最先进的串联四极杆质谱仪，其线性离子陷阱允许进行多个质谱实验，包括前体离子扫描产物离子离子扫描，中性损耗扫描，以及多次反应监测，MSN分析，直接质谱分析（MSN分析）。目前，质谱分析正在支持几项特定的研究计划，包括在转基因和嵌合小鼠中对白细胞的跨细胞生物合成的详细研究，对巨噬细胞对真菌感染的巨噬细胞反应研究中产生的脂质介质的分析以及细胞质磷脂A2的作用。生物活性活性磷脂酰丝氨酸和溶血磷脂酰丝氨酸代谢产物的结构表征与细胞凋亡相关，以及在各种肿瘤细胞与巨噬细胞和巨噬细胞细胞的一级培养的肿瘤细胞研究中产生的花生四烯酸的代谢产生的定量。这些研究的主要目的是了解脂质介质在包括癌症在内的健康和疾病中的作用，并涉及单个HPLC运行中20-30种不同代谢产物的定量测量，以获取特定生物学中蛛网酸代谢的完整图片。这也可以称为eicosanomics。还提出了研究以支持对磷脂生物化学的研究，包括在肺部感染过程中磷脂酰甘油在肺部表面活性剂中的作用，以及脂肪酸重现回到磷脂中的生物化学途径中称为“陆路”的生物化学途径。所要求的仪器具有大量多重反应监测（MRM）实验的能力，这些实验不仅可以支持二十烷组学研究，还支持与关键细胞过程以及肺部疾病有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施中，并完全致力于脂质生化研究。 公共卫生相关性：已知蛛网膜酸的代谢产物通过形成前列腺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，使用稳定的同位素稀释和串联质谱的方法是选择的方法。除此之外， 可以追求磷脂和溶物磷脂，这将揭示以前未考虑的脂质生物化学的基本生化机制。这些研究将导致我们对正常生理和病理生理过程的理解。