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

• Author / Creator

  Dawe, Margot Renee

• 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.

• Subjects / Keywords

  • Metabolomics
  • Mass-spectrometry

• Graduation date

  Fall 2011

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R3GD70

• 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.