title: Semi-targeted analysis of metabolites using capillary-flow ion chromatography coupled to high-resolution mass spectrometry.
abstract: 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.
authors: Scottish Metabolomics Facility, University of Glasgow, Glasgow, UK. karl.burgess@glasgow.ac.uk
authors: Karl
authors: K
authors: Burgess
authors: Darren
authors: D
authors: Creek
authors: Paul
authors: P
authors: Dewsbury
authors: Ken
authors: K
authors: Cook
authors: Michael P
authors: MP
authors: Barrett
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005947:Glucose / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry / Q000379:methods*; D055432:Metabolomics / Q000379:methods*; D025341:Principal Component Analysis; D011392:Proline / Q000378:metabolism; D012680:Sensitivity and Specificity; D014346:Trypanosoma brucei brucei / Q000378:metabolism
chemical_list: D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011392:Proline; D005947:Glucose