The research proposed in the analytical core falls into two part. The first involves the analytical support of three clinical projects. These projects require the analysis of two separate groups of compounds in human plasma. To assess the formation and possible role of reactive oxygen species in a variety of disease states, glutathione and glutathione disulfide will be analyzed spectrophotometrically. Eicosanoids formed from peroxidation reactions or from cyclooxygenase and lipoxygenase pathways will be analyzed by gas chromatography-mass spectrometry (GC-MS) with electron capture negative ion detection. The application of novel analytic techniques based on immunoadsorption chromatography and selected reaction monitoring (SRM) mass spectrometry should greatly enhance research capabilities in the eicosanoid field and it is the application of these techniques which will make up the second part of this analytical core. The tedious isolation and purification methods that are often required for the analysis of trace components in biological fluids can to some extent be circumvented by the use of immunoaffinity chromatography (Gaskell and Brownsey, 1983). We will apply this novel methodology to the analysis of 6-oxo PGFl alpha, thromboxane B2, and leukotriene B4. In addition, the state- of-the-art mass spectrometry that is available in our laboratory will enable us to incorporate SRM techniques into our analytical methods. The greatly improved selectivity achieved with SRM should prove particularly valuable in eicosanoid trace analyses.
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