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Permanent link (DOI): https://doi.org/10.7939/R3GD70

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Differential 15N2-/14N2-isotope Dansylhydrazine Labeling and LC-MS for Quantification of the Human Carbonyl Metabolome Open Access

Descriptions

Other title
Subject/Keyword
metabolomics
mass spectrometry
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Dawe, Margot Renee
Supervisor and department
Li, Liang (Chemistry)
Examining committee member and department
Deyholos, Michael (Biological Sciences)
Harynuk, James (Chemistry)
Department
Department of Chemistry
Specialization

Date accepted
2012-05-31T11:54:57Z
Graduation date
2011-11
Degree
Master of Science
Degree level
Master's
Abstract
The objective of this work was the design and use of paired labeling reagents that are chemically identical but isotopically different to provide a simple and robust means of quantitative mass spectrometry (MS) based metabolome profiling. Herein is presented the differential 15N2-/14N2-isotope dansylhydrazine (DH) derivatization strategy for the quantitative profiling of carbonyl compounds in the human metabolome with sensitive analysis by liquid chromatography electrospray ionization Fourier Transform ion cyclotron resonance mass spectrometry (LC-ESI FT-ICR-MS). This is a universal technique for the identification and quantification of ketones, aldehydes, keto-acids, and sugars in biofluids by the formation of a relatively hydrophobic dansylhydrazone derivative that has shown significant improvement of reversed phase LC properties and enhancement of ESI-MS signals in LC-MS. There is no observed isotope effect using reversed phase LC and the isoforms of the light- and heavy-chain labeled metabolites are co-eluted and simultaneously detected by MS, allowing for precise quantification and confident metabolite identification. Applications of this technique are presented for the analysis of the human urinary and plasma metabolomes.
Language
English
DOI
doi:10.7939/R3GD70
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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File format: pdf (Portable Document Format)
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Last modified: 2015:10:12 12:50:47-06:00
Filename: Dawe_Margot_Fall 2011.pdf
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File title: Chapter 1: Introduction
File author: Margot Dawe
Page count: 170
File language: en-CA
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