Print Email Facebook Twitter Simultaneous quantitative analysis of metabolites using ion-pair liquid chromatography-electrospray ionization mass spectrometry Title Simultaneous quantitative analysis of metabolites using ion-pair liquid chromatography-electrospray ionization mass spectrometry Author Coulier, L. Bas, R. Jespersen, S. Verheij, E. van der Werf, M.J. Hankemeier, T. TNO Kwaliteit van Leven TNO Voeding Publication year 2006 Abstract We have developed an analytical method, consisting of ion-pair liquid chromatography coupled to electrospray ionization mass spectrometry (IP-LC-ESI-MS), for the simultaneous quantitative analysis of several key classes of polar metabolites, like nucleotides, coenzyme A esters, sugar nucleotides, and sugar bisphosphates. The use of the ion-pair agent hexylamine and optimization of the pH of the mobile phases were critical parameters in obtaining good retention and peak shapes of many of the above-mentioned polar and acidic metabolites that are impossible to analyze using standard reversed-phase LC/MS. Optimum conditions were found when using a gradient from 5 mM hexylamine in water (pH 6.3) to 90% methanol/10% 10 mM ammonium acetate (pH 8.5). The IP-LC-ESI-MS method was extensively validated by determining the linearity (R2 > 0.995), sensitivity (limit of detection 0.1-1 ng), repeatability, and reproducibility (relative standard deviation <10%). The IP-LC-ESI-MS method was shown to be a useful tool for microbial metabolomics, i.e., the comprehensive quantitative analysis of metabolites in extracts of microorganisms, and for the determination of the energy charge, i.e., the cellular energy status, as an overall quality measure for the sample workup and analytical protocols. © 2006 American Chemical Society. Subject BiologyBiotechnologyChemical analysisEstersLiquid chromatographyMass spectrometryOptimizationPH effectsElectrospray ionization mass spectrometryEnergy chargeIon pair liquid chromatographyPolar metabolitesMetabolitesAcyl coenzyme AAmmonium acetateBisphosphonic acid derivativeHexylamineMethanolNucleotideSugar bisphosphonateSugar nucleotideUnclassified drugWaterAcidityAlkalinityAnalytic methodBacillus subtilisElectrospray mass spectrometryEnergy expenditureEscherichia coliIon pair chromatographyMetaboliteMetabolomicsNonhumanProcess optimizationQuantitative assayReproducibilitySamplingValidation processAminesBacillus subtilisChromatography, LiquidCoenzyme AEscherichia coliEstersHydrogen-Ion ConcentrationNucleotidesReproducibility of ResultsSpectrometry, Mass, Electrospray IonizationSugar Phosphates To reference this document use: http://resolver.tudelft.nl/uuid:df5f8f5d-ebbe-486c-8945-19401a814161 DOI https://doi.org/10.1021/ac0607616 TNO identifier 239477 ISSN 0003-2700 Source Analytical Chemistry, 78 (18), 6573-6582 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.