Simultaneous Analysis of Hydrophilic Metabolites Using Triple Quadrupole LC/MS/MS

Many primary metabolites associated with the central metabolic pathway and glycolytic system are hydrophilic compounds, which are difficult to separate using reversed phase chromatography. Ion pairing reagents may be added to the mobile phase to separate and retain polar primary metabolites on reversed phase columns. For instance, in order to measure anions, such as sugar phosphates, nucleotides and organic acids, a cationic ion pair reagent in the mobile phase can be used. In this data sheet, tributylamine was used as an ion pair reagent. It illustrates simultaneous analysis of hydrophilic metabolites using a Shimadzu UFMS triple quadrupole mass spectrometer, the LCMS-8040, with a ready-to-use "LC-MS/MS method package for Primary Metabolites".
The compound panel is illustrated below. It includes very important metabolites such as amino acids, organic acids, sugar phosphates, nucleotides and co-enzymes whose metabolic changes need to be observed. In simultaneous analysis of these metabolites, it is important to accurately reflect biological variation by preventing analyte degradation during sample preparation as much as possible. We have investigated this effect during sample preparation by comparing an intentionally degraded sample with a properly prepared sample.