Development of Isotope Labeling Liquid Chromatography Mass Spectrometry for Biofluid Metabolomics and Applications in Disease Studies

• Author / Creator

  Peng, Jun

• Metabolomics is a research field focusing on global study of all the metabolites present in a biological system (i.e., the metabolome). Metabolome analysis involves identifying and quantifying as many small molecule metabolites as possible in a biological sample. Metabolomics has attracted much attention in recent years and holds promise in application areas such as disease biomarker discovery. The objective of this thesis is to develop isotope labeling LC-MS metabolomics methods to improve the metabolome coverage and quantification precision in biofluids including human urine, mouse urine, and bronchoalveolar lavage fluids, and apply the developed metabolomics methods to study animal model of diseases including Alzheimer’s disease and asthma. We developed an offline two-dimensional LC separation strategy based on ion pairing reversed phase LC, coupled with dansylation labeling LC-MS for comprehensive metabolomic profiling of human urine. We also developed a liquid-liquid extraction method coupled with dimethylphenacyl labeling LC-MS for improved metabolomic profiling of organic acids in human urine. We developed a metabolomics workflow for comparative study of mouse model of Alzheimer’s disease and wild type, and it is demonstrated that mouse urine metabolomics could be a useful approach to study Alzheimer’s disease. We also developed an isotope labeling LC-MS method for bronchoalveolar lavage fluids and applied it to study a rat model of allergic inflammation. Metabolic pathway analysis implicates that arginine-proline metabolic pathway may be associated with allergic inflammation. Through these research activities, my thesis work has contributed to the development of isotope labeling LC-MS for metabolomics applications in disease studies and biomarker discovery.

• Subjects / Keywords

  • Metabolomics
  • Isotope labeling
  • Mass-spectrometry

• Graduation date

  Spring 2014

• Type of Item

  Thesis

• Degree

  Doctor of Philosophy

• DOI

  https://doi.org/10.7939/R3NS0M835

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.