Mass spectrometry for small molecule profiling in the Scripps Florida DMPK core

描述(由申请人提供)：本提案寻求资金购买一台配备适当软件的ThermoFisher Q-Exactive快速扫描高分辨率质谱仪，该仪器将成为药物代谢和药代动力学(DMPK)核心的一部分，专门用于研究小分子在体外和动物中的处置。这一共享工具将扩展能力，并支持一系列研究项目，这些项目源于美国国立卫生研究院资助的来自七个机构的16个项目。这些项目中的每一个与DMPK核心的参与度都很高，这体现在各自赠款中的财政承诺，以支付部分技术员工资和实验费用(16名调查人员中的13名)，或通过专门写入赠款的退款(16项中的3项)。我们提出了购买Q-Exactive质谱计系统的理由，证明了使用高分辨率质谱仪进行的新陈代谢实验所获得的信息含量显著增加。这项新技术的采用使人们能够同时从复杂样品中获取高性能的定性和定量数据。目前斯克里普斯DMPK核心中的质谱计是三重四极杆仪器，非常适合测定血浆或其他组织中的药物浓度，但缺乏高分辨率和质量精度，这对于表征未知代谢物是必不可少的。样品是在佛罗里达州斯克里普斯准备的，并被送往三家供应商的应用专家进行评估，以评估ThermoFisher Q-Exactive、ABSciex 5600 tripleTOF和Waters Synapt G2 QTOF。提供的数据显示了每个主要用户将从采用高分辨率质谱仪中获得的好处，这是目前现有设备无法实现的。文中还给出了数据，以合理地选择Q-Exactive而不是其他优秀的系统。受益于共享仪器的大部分赠款是探测器优化、转化科学和药物发现赠款。在优化阶段，阐明单个化合物的代谢负债的能力显著增加，将提高项目生产率。化合物优化是一个迭代的过程，在早期阶段了解分子的药物新陈代谢和药代动力学特性，可以优化整个分子，而不仅仅是生化或细胞效力。这台共享仪器将安装在佛罗里达州斯克里普斯质谱学实验室，并将由一组经验丰富的质谱学家、PI和两名高级研究技术人员进行维护。几名博士后研究员将接受该仪器的培训或继续他们的培训。

DESCRIPTION (provided by applicant): This proposal seeks funding to purchase a ThermoFisher Q-Exactive rapid scanning high-resolution mass spectrometer with appropriate software that will become part of the drug metabolism and pharmacokinetic (DMPK) core dedicated to the study of small molecule disposition in vitro and in animals. This shared instrument will expand capabilities and support a range of research projects originating from sixteen NIH funded projects from seven institutions. The level of engagement for each of these projects with the DMPK core is high as demonstrated by the financial commitments in the respective grants to cover portions of technician salary and experimental expenses (13 of 16 investigators) or through chargebacks that are specifically written into grants (3 of 16). We present justification for the purchase of a Q-Exactive mass spectrometer system by demonstrating significant increases to the information content obtained from metabolism experiments performed using high resolution mass spectrometry. Incorporation of this new technology allows for the simultaneous acquisition of high-performance qualitative and quantitative data from complex samples. The current mass spectrometers in the Scripps DMPK core are triple-quadropole instruments ideally suited for determination of drug concentrations in plasma or other tissues, but lacking high resolution and mass accuracy which are essential for the characterization of unknown metabolites. Samples were prepared as Scripps Florida and sent to application specialists at three vendors for evaluation of the ThermoFisher Q-Exactive, the ABSciex 5600 tripleTOF, and the Waters Synapt G2 QTOF. The presented data demonstrates the benefits each major users will derive from the incorporation of high resolution mass spectrometry that are not currently possible with existing equipment. Data is also presented to rationalize the choice of the Q-Exactive over the other excellent systems. The majority of the grants that benefit from the shared instrument are probe optimization, translational science, and drug discovery grants. Significant increases in capacity to elucidate metabolic liabilities of individual compounds during the optimization phase will increase project productivity. Compound optimization is an iterative process and understanding the drug metabolism and pharmacokinetic properties of molecules at an early stage allows the whole molecule to be optimized instead of just the biochemical or cellular potency. This shared instrument will be installed in the Scripps Florida Mass Spectrometry Laboratory and will be maintained by a group of experienced mass spectroscopists, the PI and two senior research technicians. Several Post Doctoral fellows will be trained or continue their training on this instrument.