CORE--MASS SPECTROMETRY

近10年来，核心质谱仪继续为各个项目提供分析支持。质谱法已成功地用于各种研究，并已被证明是一种有价值的和适当的技术，在本申请中提出的研究类型。质谱法是正在进行的研究的一个重要方面，将被所有计划项目研究人员用于两个主要目的：（a）定量分析花生四烯酸和一氧化碳的PO 450衍生代谢物，和（B）定性（结构）分析。我们在实验室中开发了基于同位素稀释GC/MS的CO、20-HETE和其他亚末端HETE、环氧二十碳三烯酸（ECO 2）的异构体以及ω-羟基化的胡萝卜素的定量分析。由于来自各个研究者的所有样品都是由这个具有长期质谱经验的专用核心设施进行分析的，因此可以以统一的方式获得定量数据，具有高质量控制和最佳成本。在整个研究期间，在相同条件下分析样品。此外，我们还开发了一套样品处理和分析系统，可有效利用仪器和工作人员的时间，降低运营成本，同时及时提供准确的结果。利用质谱分析的各个方面是本项目建议书的一个重要特点。这源于认识到质谱分析提供了足够的灵敏度、特异性和选择性，这是检测和表征通常在生物材料中发现的亚皮摩尔量的类二十烷酸所需的。特别是，通过细胞色素P450途径形成的代谢物或花生四烯酸只能使用GC/MS在亚皮摩尔水平下准确和精确地测量，因为目前没有其他具有可比灵敏度和特异性的技术可用于其定量。核心功能的一个新方面是开发一种精确的方法，用于定量分析源自生物材料的CO。使用这些方法，核心将支持计划项目研究人员努力回答有关类花生酸在高血压机制中的作用的基本问题。

The Core Mass Spectrometry has continued to provide analytical support for the individual program projects for almost 10 years. Mass spectrometry has been successful used in various studies and has proven to be a valuable and adequate technique for the type of studies proposed in this application. Mass spectrometry is an important aspect of the ongoing research and will be utilized by all program project investigators essentially for two general purposes: (a) quantitative analyses of PO450- derived metabolites of arachidonic acid and carbon monoxide, and (b) qualitative (structural) analyses. We have developed in our laboratory quantitative assays based on isotopic dilution GC/MS for CO, 20-HETE and other subterminal HETEs, isomers of epoxyeicosatrienoic acids (EETs) and also for omega-hydroxylated prostaglandins. Because all samples from individual investigators are analyzed by this dedicated core facility with long experience in mass spectrometry, the quantitative data are obtained in a uniform manner, with high quality control and at optimal cost. Samples are analyzed at the same conditions through the study period. Additionally, we have developed a system for sample handling and analysis that utilizes the instrument and staff time efficiently and reduces operational costs while delivering accurate results in a timely manner. Utilization of various aspects of mass spectrometry is an important feature of this program project proposal. This stems from the recognition that mass spectrometric analysis provide adequate sensitivity, specificity, and selectively required for detection and characterization of sub-picomolar quantities of eicosanoids typically found in the biological material. In particular, metabolites or arachidonic acid formed via cytochrome P450 pathway can only e measured accurately and precisely using GC/MS at subpicomolar levels since no other technique of comparable sensitivity and specificity is currently available for their quantitation. A novel aspect of the Core function is the development of an accurate method for quantitative analysis of CO originating from biological material. Using these methodologies, the Core will support Program Project Investigators in their efforts to answer fundamental questions regarding the role of eicosanoids in the mechanisms of hypertension.