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

购买 AB Sciex Qtrap 6500 LC/MS/MS 系统

• 批准号: 8824667
• 负责人: LAWRENCE J. MARNETT
• 金额: $ 44.96万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8824667
• 关键词: 2-arachidonylglycerol; 7-dehydrocholesterol; Accounting; Adoption; Advisory Committees; Biochemistry; Biological Assay; Biology; Chemicals; Chemistry; Complex; Data; Detection; Endocannabinoids; Funding; Guidelines; Housing; Hydrolysis; Inflammation; Lipids; Maintenance; Mass Spectrum Analysis; Metabolic; Metabolism; Monitor; Pain; Pharmaceutical Chemistry; Request for Applications; Research; Research Personnel; Research Support; Sampling; Scanning; Staging; Sterols; Stress; Surveys; System; Therapeutic; Training; Translations; United States National Institutes of Health; Work; anandamide; cholesterol biosynthesis; high throughput screening; in vivo; in vivo Model; instrument; interest; liquid chromatography mass spectrometry; metabolomics; novel; oxidation; research study; response

 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.

 描述（由申请人提供）：本申请申请要求资金购买专用的AB Sciex Qtrap 6500 LC-MS/MS系统，以允许对复杂样品中的脂质代谢物进行高灵敏度分析，并对范德比尔特高通量筛选设施（VHTS）生成的样品进行快速自动分析。该仪器将置于范德比尔特质谱研究中心（MSRC）的质谱核心内并由其操作。它将支持一组主要用户的研究，包括Lawrence Marnett，Sachin Patel和Roger Colbran，他们研究内源性大麻素代谢对疼痛，炎症和压力的反应的化学和生物学。他们的工作需要量化内源性大麻素，2-花生四烯酸甘油（2-AG）和花生四烯酸乙醇酰胺（AEA），以及它们通过水解或氧化形成的代谢物。主要用户内德·波特研究7-脱氢胆固醇的化学和生物化学，7-脱氢胆固醇是胆固醇生物合成的中间体，极易氧化。他的工作需要 这种甾醇及其众多氧化产物的定量。所有这些研究者采用体内模型，其提供有限量的材料用于定量低丰度分析物。范德比尔特目前可用的仪器不够灵敏，无法检测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，可以采用各种串联MS实验作为计划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，它结合了早期代谢测定，以评估新型生物活性化合物的潜在体内稳定性。这些测定以高通量形式（96孔和384孔板）进行，其提供了大量用于分析的含有有限量代谢物的样品。这些研究对于越来越多的NIH资助的研究人员的药物化学工作至关重要，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小组对高灵敏度质谱法的有限访问已经排除了对化合物翻译至关重要的代谢测定的常规采用。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。主要用户组将占仪器使用量的约76%，其余部分将提供给需要高灵敏度分析的其他范德比尔特研究者。质谱核心咨询委员会将监测仪器使用情况，并制定非主要用户使用标准。