A novel ion pairing LC/MS metabolomics protocol for study of a variety of biologically relevant polar metabolites.
We report a method of ion-pairing liquid chromatography coupled to mass spectrometry (IP-LC-MS) that we have developed for the sensitive detection and quantification of a variety of biologically relevant polar molecules. We use the ion-pairing agent diamyl ammonium to improve chromatographic resolution of polar compounds, such as nucleotide cofactors, sugar phosphates, and organic acids, that are generally poorly retained by conventional reverse phase chromatographic methods. This method showed good linearity (average R value of 0.996) and reproducibility (generally RSD values <10%). We demonstrate the utility of this method by investigating the metabolomic signature of three distinct biological systems: the metabolic response to lack of superoxide dismutase activity and to paraquat induced oxidative stress, and the metabolic profiles of four different Drosophila species.
['Knee|Jose M|JM|', 'Rzezniczak|Teresa Z|TZ|', 'Barsch|Aiko|A|', 'Guo|Kevin Z|KZ|', 'Merritt|Thomas J S|TJ|']
D000085:Acetates D000818:Animals D002853:Chromatography, Liquid D004330:Drosophila D005260:Female D057927:Hydrophobic and Hydrophilic Interactions D057230:Limit of Detection D016014:Linear Models D008297:Male D013058:Mass Spectrometry D055442:Metabolome D055432:Metabolomics D008956:Models, Chemical D015203:Reproducibility of Results
Polar metabolites Diamyl ammonium (DAA) Drosophila metabolomics IP-LC–MS Ion pairing chromatography