Author

Tao Sun

Defense Date

2-28-2012

Graduation Date

2012

Availability

Immediate Access

Submission Type

dissertation

Degree Name

PhD

Department

Chemistry and Biochemistry

School

Bayer School of Natural and Environmental Sciences

Committee Chair

Jana Patton-Vogt

Committee Member

Partha Basu

Committee Member

Rita M. Mihailescu

Committee Member

Stephanie J. Wetzel

Keywords

High performance liquid chormatography, Lipidomics, Mass spectrometry, Metabolics, Microfluidics, Yeast

Abstract

Lipidomics and metabolomics are powerful tools for the examination of cellular metabolism and physiology. Methods for lipid analysis need to be developed that begin with small samples and do not overly dilute or disperse the sample in the separation process. Microchips provide a platform for interfacing lysis of small cell populations with on-chip solid phase extraction for isolating lipid samples to generate high quality mass spectra from very small samples. Chapter 1 of this dissertation presents a novel method for small scale lipidomics of bacterial cells by microchip based extraction coupled with untargeted profiling of glycerophospholipids using nanoelectrospray ionization mass spectrometry. Chapter 2 and 3 focus on the development of LC-MS/MS methods to study biological pathways. In Chapter 2, I describe a method for analysis of the phospholipids metabolite, GroPIns, in the medium of the pathogenic yeast Candida albicans. This method was applied to aid in the characterization of the GroPIns transport protein, Git1, in C. albicans. Chapter 3 extends the studies of part two and describes an efficient method based on HILIC-MS/MS for the separation and quantification of five lipid-related extracellular metabolites in yeast Saccharomyces cerevisiae. This newly developed methodology was successfully applied to determine the extracellualr levels of glycerophosphoinositol, glycerophosphocholine, glycerol 3-phosphate, inositol and choline in wild type and mutant strains.

Format

PDF

Language

English

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