Acquisition of an AB Sciex Qtrap 6500 LC/MS/MS System

 描述(由申请人提供)：本申请申请资金用于购买专门的AB Sciex QTrap 6500 LC-MS/MS系统，以便对复杂样品中的脂类代谢物进行高灵敏度分析，并能够快速、自动化地分析Vanderbilt High Throughput Screen Equipment(VHTS)产生的样品。该仪器将安装在范德比尔特质谱学研究中心(MSRC)的质谱学核心中并由其操作。它将支持一组主要使用者的研究，包括劳伦斯·马内特、萨钦·帕特尔和罗杰·科尔布兰，他们研究内源性大麻素代谢对疼痛、炎症和压力的化学和生物学反应。他们的工作需要对内源性大麻素、2-花生四烯基甘油(2-AG)和花生四烯基乙醇胺(AEA)及其通过水解或氧化形成的代谢物进行量化。主要使用者内德·波特研究7-脱氢胆固醇的化学和生化，7-脱氢胆固醇是胆固醇生物合成的中间体，极易被氧化。他的工作需要 对这种甾醇及其众多氧化产物进行定量。所有这些研究人员都采用了体内模型，这些模型为量化低丰度分析物提供了有限的材料。范德比尔特目前可用的仪器不够灵敏，无法检测到许多感兴趣的分析物。此外，主要用户需要快速的数据依赖扫描，可以使用各种串联MS实验作为计划中的代谢组学实验的调查扫描。QTrap 6500的第五个主要用户将是VHTS，它已经纳入了早期代谢分析，以评估新的生物活性化合物的潜在体内稳定性。这些分析是以高通量格式(96孔板和384孔板)进行的，它提供了大量样品供分析，但代谢物的含量有限。这些研究对越来越多的NIH资助的研究人员的药物化学努力至关重要，他们需要这些研究来最大限度地提高生物活性化合物的体内稳定性，以用作机制和治疗概念验证的化学探针。VHTS小组获得高灵敏度质谱仪的机会有限，这排除了对化合物翻译至关重要的代谢分析的常规采用。QTrap 6500 LC-MS/MS将遵守MSRC的操作指南，MSRC还将负责其维护和用户培训。主要用户组将占该仪器使用的大约76%，其余部分提供给其他需要高灵敏度分析的范德比尔特调查人员。质谱学核心咨询委员会将监测仪器的使用情况，并为非主要用户的使用制定标准。

 DESCRIPTION (provided by applicant): This application requests funds to purchase a dedicated AB Sciex Qtrap 6500 LC-MS/MS system to allow high-sensitivity analysis of lipid metabolites in complex samples, as well as to enable rapid, automated analysis of samples generated by the Vanderbilt High Throughput Screening Facility (VHTS). The instrument will be housed within and operated by the Mass Spectrometry Core of the Vanderbilt Mass Spectrometry Research Center (MSRC). It will support the research of a group of major users, including Lawrence Marnett, Sachin Patel, and Roger Colbran, who study the chemistry and biology of endocannabinoid metabolism in response to pain, inflammation, and stress. Their work requires quantification of the endocannabinoids, 2-arachidonoylglycerol (2-AG) and arachidonoylethanolamide (AEA), and their metabolites formed by hydrolysis or oxygenation. Major user Ned Porter studies the chemistry and biochemistry of 7-dehydrocholesterol, an intermediate in cholesterol biosynthesis that is highly susceptible to oxidation. His work requires quantification of this sterol and its numerous oxidation products. All of these investigators employ in vivo models that provide limited amounts of material for quantification of low abundance analytes. The instruments currently available at Vanderbilt are not sensitive enough to enable detection of many of the analytes of interest. Furthermore, the major users require rapid data-dependent scanning that can employ a variety of tandem MS experiments as the survey scan for planned metabolomics experiments. The fifth major user of the QTrap 6500 will be the VHTS, which has incorporated early-stage metabolic assays to assess the potential in vivo stability of novel bioactive compounds. These assays are done in high-throughput format (96-well and 384-well plates), which provides large numbers of samples for analysis that contain limited amounts of metabolites. These studies are critical to the medicinal chemistry efforts of a growing number of NIH-funded investigators, who need them to maximize the in vivo stability of bioactive compounds for use as chemical probes of mechanism and therapeutic proofs-of-concept. The limited access of the VHTS group to high sensitivity mass spectrometry has precluded routine adoption of the metabolic assays so critical for compound translation. The QTrap 6500 LC-MS/MS will be subject to the operating guidelines of the MSRC, which will also take responsibility for its maintenance and user training. The Major user group will account for approximately 76% of the use of the instrument, with the remainder made available to other Vanderbilt investigators requiring high sensitivity analysis. The Mass Spectrometry Core Advisory Committee will monitor instrument use and establish criteria for access by non-major users.