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Permanent link (DOI): https://doi.org/10.7939/R3N873B8R

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Development and Application of Isotope Labeling Method and Data Processing Program for Liquid Chromatography Mass Spectrometry Based Metabolome Profiling Open Access

Descriptions

Other title
Subject/Keyword
Ginseng Root
Goji Tea
Plant Fibers
Arabidopsis Thaliana
IsoMS
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Tseng, Chiao-Li
Supervisor and department
Li, Liang (Chemistry)
Examining committee member and department
Harynuk, James (Chemistry)
McCreery, Richard L. (Chemistry)
Gabryelski, Wojciech (Chemistry, University of Guelph)
Clive, Derrick L. J. (Chemistry)
Department
Department of Chemistry
Specialization

Date accepted
2015-02-03T14:17:59Z
Graduation date
2015-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
The objective of this thesis focuses on establishing a software tool (IsoMS) used in the quantification and qualification of metabolites in complex biological samples, developing robust methods to extract metabolites from different plant samples, and applying a differential 13C-/12C-isotope dansylation labeling technique with liquid chromatography-mass spectrometry (LC-MS) for the analysis of amine- and phenol-containing metabolites in plant and human urine samples. This approach, a differential 13C-/12C-isotope dansylation labeling method combined with LC-MS, has successfully been applied to the relative quantification, peak pair identification, and putative metabolite identification in a study of metabolic profiling. First of all, IsoM has been developed to handle the raw LC-MS data generated by a chemical isotope labeling or isotope coded derivatization (ICD) metabolomics platform by peak picking, peak pairing, peak-pair filtering and peak-pair intensity ratio calculation. Secondly, a protocol for rapid extraction, derivatization and detection of amine- and phenol-containing metabolites in mg-scale samples of flax (Linum usitiatissimum) bast fibers has been developed and can typically be completed in under 13 hours for up to 8 samples. Thirdly, the development of a robust metabolite extraction method from ginseng roots tailored to the analysis of amine- and phenol-containing metabolites and the use of the dansylation labeling method to gauge the detectability of the LC-MS technique for the ginseng metabolome has been used on the application of spatial distribution of metabolites in ginseng roots. Fourthly, a microwave-assisted extraction method of Arabidopsis thaliana has been optimized and used prior to a differential 13C-/12C-isotope dansylation labeling technique with LC-FT-MS for the study of metabolic profiling of different TIFY gene expression plants under methyl jasmonate treatment. Finally, a highly sensitive dansylation isotope labeling LC-MS method was used to determine the urine metabolomes before and after a moderate amount of drinking Goji tea, considered as a phyto-nutritional supplement drink. The result clearly showed that the consumption of Goji tea does not affect the urine metabolome significantly for the studies of the short term (<3 hr) and longer term (12 hr) effects of drinking Goji tea.
Language
English
DOI
doi:10.7939/R3N873B8R
Rights
This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for the purpose of private, scholarly or scientific research. 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.
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