ANALYTICAL CORE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 分析核心：分析核心的目的是为参与研究的研究人员提供测量各种生物分子的最新方法。 该目标已经通过使用两种互补技术实现：HPLC串联质谱法和基于多重荧光（“Luminex”）的测定。 核心由安德鲁·莫里斯博士执导，内科学和分子与细胞生物化学教授。 Manjula Sunkara，MS，研究助理，负责核心仪器的日常操作。 Sunkara女士拥有分析化学背景，并在一家制药研究公司获得了生物医学质谱分析方面的额外经验。 Sunkara女士完全得到了COBRE奖的支持。 核心的运营费用由参与的研究人员分摊，部分由研究副总裁提供的资金支持，以促进其他非COBRE附属研究人员使用该仪器。 利用先前获得的NCRR共享仪器资助（1 S10 RR 024598 -01，Andrew J. Morris，PI）提供的资金，肯塔基州大学医学院的一个互动研究小组最近购买了一台应用生物系统（ABI）/MDS Sciex 4000 Q-Trap混合三重四极杆线性离子阱质谱仪。 该仪器配备有ABI Turbo V电喷雾电离离子（ESI）源，并与配备有柱温箱和自动进样器的Shimadzu多泵HPLC系统连接。 运行ABI Analyst软件的计算机工作站用于仪器控制以及数据采集和处理。 该仪器于6个月前投入使用，在此期间，我们建立了多反应监测模式HPLC串联质谱分析，用于分析和定量广泛和扩展范围的脂质和脂质相关分子，包括甘油和鞘磷脂、脂肪酸、类花生酸、类异戊二烯和甾醇。 使用该仪器获得的数据已发表在《生物化学和分子药理学杂志》上，其他报告和赠款申请来自使用目前正在审查的仪器进行的实验。 核心还包含Luminex（BioRad）200多重荧光分析仪。 我们提供校准、实验设计方面的协助，并与适当的供应商合作，开发针对参与研究者需求量身定制的分析物集合“面板”。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ANALYTICAL CORE: The purpose of the Analytical Core is to provide state of the art methodologies for measurement of a broad range of biomolecules to the participating investigators. This goal has been accomplished by use of two complementary technologies: HPLC Tandem mass spectrometry and multiplex fluorescence ("Luminex")-based assays. The core is directed by Andrew Morris, Ph.D., Professor of Internal Medicine and Molecular and Cellular Biochemistry. Manjula Sunkara, MS, Research Associate, is responsible for day to day operation of the core instrumentation. Ms Sunkara has a background in analytical chemistry with additional experience in biomedical mass spectrometry gained with a pharmaceutical research company. Ms Sunkara is supported entirely by the COBRE award. Operating expenses of the core are cost shared by the participating investigators and supported in part by funds provided by the Vice President for Research to facilitate use of the instrument by other non-COBRE affiliated researchers. Using funds provided by a previously awarded NCRR Shared instrumentation grant (1S10RR024598-01, Andrew J. Morris, PI) an interactive group of researchers at the University of Kentucky College of Medicine recently purchased an Applied Biosystems (ABI)/MDS Sciex 4000 Q-Trap hybrid triple quadrupole linear ion trap mass spectrometer. The instrument is equipped with an ABI Turbo V electrospray ionization ion (ESI) source and interfaced with a Shimadzu multi pump HPLC system equipped with a column oven and autosampler. A computer workstation running ABI Analyst Software is used for instrument control and data acquisition and processing. The instrument was put into service about 6 months ago and in the intervening time we have set up multiple reaction monitoring mode HPLC tandem mass spectrometry assays for analysis and quantitation of a broad and expanding range of lipids and lipid-related molecules which include glycero and sphingo phospholipids, fatty acids, eicosanoids, isoprenoids and sterols. Data obtained using this instrument have been published in the Journal of Biological Chemistry and Molecular Pharmacology with other reports and grant applications arising from experiments conducted using the instrument currently under review. The core also contains a Luminex (BioRad) 200 multiplex fluorescence analyzer. We provide assistance with calibration, experimental design and are working with the appropriate vendors to develop "panels" for collections of analytes that are tailored to the needs of the participating investigators.