文献汇报辉.pptVIP

Development of an online SPE–LC–MS-based assayusing endogenous substrate for investigation of solubleepoxide hydrolase inhibitors 报告人：谢辉 2011年6月29日 Background 1.Therapeutic potency of sEHI sEH inhibitors are a promising new class of potential drugs for treatment of a variety of diseases, for example inflammation, hypertension, and pain. 2. New assays are needed Analytical techniques are needed to identify active compounds and quantitatively measure their potencies. 3. Development of new assay electrospray ionization tandem mass spectrometry (ESI–MS–MS) (电喷雾离子阱质谱) Merits Compared to fluorescence，this method enables quantitative evaluation of potent sEHIs with picomolar potencies . (13 different inhibitors / individual compounds) So this assay is a promising method！ Procedure 1. Selection of substrates Natural substrates : 14(15)-EpETrE Internal standards : 10(11)-EpHep Baculovirus-expressed human sEH 2. Selection of sEHI Urea derivatives synthesized in their laboratory Baculovirus-expressed human sEH Method MS (mass spectrometry) —measurement of enzyme activity and inhibition LC–ESI（Liquid chromatography electrospray） — to monitor conversion of 14(15)-EpETrE to 14,15-DiHETrE (The method is a new use for the rapid determination of the potency of sEHI.) SPE ( solid-phase extraction ) — to ensure sensitive and reliable determination of small amounts of product formed in a difficult matrix. method applied here — short, narrow columns filled with large particles (50–60 μm) —Turbulent-flow chromatography (TFC ). 3. Three steps 1.Preparation of stock and standard solutions 2. Instrumental setup 3. sEH inhibition assay Results Summary A new LC–MS-based natural substrate approach for potency measurement of sEHI has been developed. This new method enables reliable measurement of the potency of sEHI with high precision in less time. 固相萃取的简要过程 1. 一个样品包括分离物和干扰物通过吸附剂 2. 吸附剂选择性的保留分离物和一些干扰物其