The Chemistry Core provides chemical services and logistical support to the constituent laboratories of the program project. Specifically, it has three main responsibilities: (1) Prepare labeled and unlabeled eicosanoids, standards, metabolite analogous, and inhibitors in sufficient quantities for biological studies, such as assay standards, for structure elucidations and for methodology development. In light of the limited availability of most eicosanoids from natural sources, the synthetic and analytical resources of the Core are essential for the successful conduct of the program project. All synthases are patterned on existing synthetic routes developed in Project VI or adapted from the literature. Generally, it is necessary to make modifications to the syntheses to accommodate the increased scale of the reactions or to permit the incorporation of labeled atoms. For the latter, commercially labeled fatty acids are convenient precursors or labeled regents such as [2H, 3H, 18O]-H2O,[14C]-NaCN, [2H, 3H]-H2 gas, [2H, 3H]- LiAIH/4/NaBH4 are used. Appropriate analytical techniques, e.g., capillary GLC, HPLC, NMR, and mass spectrometry, ensure product identify, level of purity, and isotopic composition (if present). (2) Develop and validate analytical techniques for the identification, stereochemical confirmation, and quantitation of renal eicosanoids. Metabolites of the P450 arachidonate pathways are good candidates of initial immunoassay development since they are novel eicosanoids with relevance to kidney function and because they lack a chromophore suitable for UV detection and quantification. Polyclonal antibodies will be raised using rabbits and mice will serve for the production of monoclonal antibodies. Additional, electron capture, fluorescence, and chiral chromatographic protocols will be established to improve detection sensitivity, attain better resolution, and to help elucidate stereochemistry. (3) Prove for the storage, quality assurance, packaging, and distribution of eicosanoids, inhibitors, analogs, reagents, and antibodies to collaborating laboratories.
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