The purpose of this R21 application is to develop an automated, chip-based metabolomic analysis for point of-practice applications. Intracellular metabolite measurements in current forms are tedious, inaccurate, and expensive. However, their importance has grown significantly as various high-throughput technologies for genomics, transcriptomics, and proteomics have been developed. The measurement of metabolite profiles becomes the next challenge before integrating various """"""""omic"""""""" data to explain complex physiology at the systems level. Typically, intracellular metabolite measurements require extensive operations for quenching the metabolism and extracting metabolites from the cell. Because of the relatively fast turnover of metabolites, the quenching and extraction efficiency become major hurdles in their measurements. In addition, metabolite measurements typically require liquid chromatography/mass spectrometry (LC/MS) or gas chromatography (GC)/MS, which may not be readily available in each laboratory, and the cost of instrumentation, is often prohibitive. This application proposes to solve these problems by developing a low cost, automated, chip-based sample preparation and HPLC system, which can be used in common laboratories next to the point-of-practice (either experimental or clinical). The HPLC output is fractionated by an on-chip fractionator, which is physically separated from, but ready to interface with, MS/MS in other laboratories. In this scheme, sample preparation is much more accurate and convenient because of automation; metabolite separation is more efficient because tandem HPLC can be build on chip; cost of measurement is significantly reduced because of the low cost of on-chip HPLC and outsourcing of the MS/MS operation to MS specialists.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
5R33DK070328-02
Application #
7032352
Study Section
Special Emphasis Panel (ZRG1-EMNR-J (50))
Program Officer
Castle, Arthur
Project Start
2005-04-01
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2008-03-31
Support Year
2
Fiscal Year
2006
Total Cost
$1,001,393
Indirect Cost
Name
University of California Los Angeles
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
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Lin, James A; Watanabe, Junji; Rozengurt, Nora et al. (2007) Atherogenic diet causes lethal ileo-ceco-colitis in cyclooxygenase-2 deficient mice. Prostaglandins Other Lipid Mediat 84:98-107
Narasimha, Ajay; Watanabe, Junji; Lin, James A et al. (2007) A novel anti-atherogenic role for COX-2--potential mechanism for the cardiovascular side effects of COX-2 inhibitors. Prostaglandins Other Lipid Mediat 84:24-33