CORE--MASS SPECTROMETRY

近10年来，Core质谱仪一直为各个项目提供分析支持。质谱法已经成功地应用于各种研究中，并已被证明是一种有价值的和适当的技术，用于本应用中提出的研究类型。质谱法是正在进行的研究的一个重要方面，将被所有项目研究者用于两个一般目的：(a)定量分析花生四烯酸和一氧化碳的PO450衍生代谢物，以及(b)定性（结构）分析。我们在实验室开发了基于同位素稀释GC/MS的定量分析，用于CO， 20-HETE和其他亚末端hete，环氧二十碳三烯酸（EETs）的异构体以及omega-羟基化前列腺素。由于来自个别研究者的所有样品都由这个具有长期质谱分析经验的专用核心设施进行分析，因此定量数据以统一的方式获得，具有高质量控制和最佳成本。在整个研究期间，样品在相同的条件下进行分析。此外，我们还开发了一个样品处理和分析系统，有效地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。利用质谱法的各个方面是本方案的一个重要特点。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，用于检测和表征亚皮摩尔量的类二十烷类化合物，这些类二十烷类化合物通常存在于生物材料中。特别是，代谢物或通过细胞色素P450途径形成的花生四烯酸只能在亚皮摩尔水平上使用GC/MS准确和精确地测量，因为目前没有其他可用于定量的灵敏度和特异性相当的技术。核心功能的一个新方面是开发了一种精确的方法来定量分析来自生物材料的CO。使用这些方法，核心将支持项目研究者努力回答关于类二十烷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.