Premium
Semi‐targeted analysis of metabolites using capillary‐flow ion chromatography coupled to high‐resolution mass spectrometry
Author(s) -
Burgess Karl,
Creek Darren,
Dewsbury Paul,
Cook Ken,
Barrett Michael P.
Publication year - 2011
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.5247
Subject(s) - chemistry , hydrophilic interaction chromatography , chromatography , metabolomics , mass spectrometry , metabolite , liquid chromatography–mass spectrometry , metabolome , high performance liquid chromatography , biochemistry
This work describes a novel application of capillary‐flow ion chromatography mass spectrometry for metabolomic analysis, and comparison of the technique to octadecyl silica and hydrophilic interaction chromatography (HILIC)‐based mass spectrometry. While liquid chromatography/mass spectrometry (LC/MS) is rapidly becoming the standard technique for metabolomic analysis, metabolomic samples are extremely heterogeneous, leading to a requirement for multiple methods of analysis and separation techniques to perform a ‘global’ metabolomic analysis. While C18 is suitable for hydrophobic metabolites and has been used extensively in pharmaceutical drug metabolism studies, HILIC is, in general, efficient at separating polar metabolites. Phosphorylated species and organic acids are challenging to analyse and effectively quantitate on both systems. There is therefore a requirement for an MS‐compatible analytical technique that can separate negatively charged compounds, such as ion‐exchange chromatography. Evaluation of capillary flow ion chromatography with electrolytic suppression was performed on a library of metabolite standards and was shown to effectively separate organic acids and sugar di‐ and tri‐phosphates. Limits of detection for these compounds range from 0.01 to 100 pmol on‐column. Application of capillary ion chromatography to a comparative analysis of energy metabolism in procyclic forms of the parasitic protozoan Trypanosoma brucei where cells were grown on glucose or proline as a carbon source was demonstrated to be more effective than HILIC for detection of the organic acids that comprise glucose central metabolism and the tricarboxylic acid (TCA) cycle. Copyright © 2011 John Wiley & Sons, Ltd.