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Towards identifying the full metabolome of the protozoan parasite Leishmania by using high resolution liquid chromatography mass spectrometryWang L1, Watson DG1, Blackburn G1, Williams RAM3, Westrop GD1, Smith TK2, Coombs GH11 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, UK2 Schools of Biology & Chemistry, The University of St. Andrews, St. Andrews, UK3 institute of Biomedical and Environmental Health Research, University of the West of Scotland, Paisley, UKLeishmania is a protozoan parasite of humans and other mammals transmitted by the sandfly. Extensive knowledge is available on the genomes and transcriptomes of the parasite species, and these have facilitated predictions on the metabolic pathways operating. Experimental investigations over many decades have provided much information on the metabolites in the parasite. However, these have largely been focused studies rather than global profiling. We have now taken advantage of the advent on improved technologies for analysing small metabolites in biological samples to provide robust data on the whole spectrum of metabolites that comprise the metabolome in this parasite. Initially we have focussed on the promastigote stage of Leishmania major and applied methods modified from those that we used to investigate the mechanism of drug resistance in Leishmania and a reversed phase chromatography method to analyse the non-polar metabolites . We have confirmed identities of metabolites by running pure compounds as standards and MS2 for compounds where standards were not available. We have also quantified less expected metabolites via spiking with standards for these compounds over a calibration range. The analysis so far has firmly identified >400 polar and non-polar metabolites. The analysis will continue by applying IC methodologies to establish the identity of metabolites less well separated using our current HILIC methods. We expect that the analysis will provide for the parasitology community robust information on a large percentage of the metabolome together with key information on how the individual metabolites can be confirmed. Further sample manipulations will be carried out in order to extend metabolite coverage within this significant group of parasites.OverviewMetabolomic profiling of the parasite Leishmania was carried out using hydrophilic interaction chromatography with high resolution mass spectrometry detection. The analysis so far has firmly identified >400 polar and non-polar metabolites by comparison with 200 authentic standards and by using MS2 experiments to aid in identification of compounds where no standards were available. Metabolites of particular interest within the parasite were quantified by spiking with standards for the compounds over a calibration range.