This report demonstrates the proof of concept to determine eight nucleotides and six organic acid metabolites by anion-exchange chromatography with suppressed conductivity detection. The metabolites were separated on the 2 mm i.d. Thermo Scientific Dionex IonPac AS11-HC-4um column, using electrolytically generated KOH gradient. These experiments were facilitated by the Thermo Scientific Dionex ICS-6000 HPIC system. Compare these results against those using methanol organic modifier (another record in AppsLab and in the attached report).
This report demonstrates the proof of concept to determine eight nucleotides and six organic acid metabolites by anion-exchange chromatography with suppressed conductivity detection. The metabolites were separated on the 2 mm i.d. Thermo Scientific Dionex IonPac AS11-HC-4um column, using electrolytically generated KOH gradient with methanol as the organic modifier to resolve malate and succinate. These experiments were facilitated by the Thermo Scientific Dionex ICS-6000 HPIC system.
A platform using Dionex ion chromatography (IC) coupled to triple quadrupole mass spectrometry (MS) was developed to measure various metabolites in Brassica napus seeds. The IC tandem mass spectrometry (IC-MS/MS) technique provides a good tool to identify and quantify metabolites in a complex matrix.
Here, the author describes an ion-exchange system that provides good separation of the polar metabolities with on-line desalting to allow MS detection. This technique will allow studies of key metabolites that do not separate well on traditional reversed-phase columns. Some ionic metabolites may be positional isomers adjacent to each other in a synthetic pathway and as such, are isobaric. In such cases, MS is not sufficient to differentiate the two compounds. There is clearly a need for adequate separation techniques for isomeric metabolites.
