Evaluation of Derivatization Reaction Conditions for Metabolite Labelling Using Differential Isotopic Labelling for Biological Samples Analyzed by Liquid Chromatography Mass Spectrometry

• Author / Creator

  Curle, Jared P

• The objective of this work was to assess the impact of reaction conditions for isotopic labeling of metabolites prior to analysis using liquid chromatography mass spectrometry (LC-MS). Differential isotope 12C/13C dansyl chloride was used to label metabolites containing either primary amines, secondary amines, or phenolic functional groups and those were determined by both liquid chromatography electrospray ionization Fourier transform ion cyclotron resonance MS (LC-ESI-FTICR-MS) and ultra-performance liquid chromatography with ultraviolet absorbance detection (UPLC-UV). Using UPLC-UV, metabolite standards labeled under varying reaction conditions were quantitatively determined for comparison. Compared with conventional heating, microwave assisted heating (MAH) methods demonstrated better labeled product yields. In addition ambient temperature reactions were found provide the best labelling without using MAH. It was found that a reaction temperature of 40°C provided the best results based on the number of peak pairs found. However other reaction methods provided unique peak pairs not found in the other methods.

• Subjects / Keywords

  • Metabolomics
  • Isotopic Labeling
  • Dansylation

• Graduation date

  Fall 2013

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R31X1V

• 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.