ANALYTICAL CORE

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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.