Separation of Compounds in TCA Cycle on Zodiac HST RP Column

The citric acid cycle (also known as the tricarboxylic acid cycle or Krebs cycle) is a central metabolic pathway through which aerobic organisms produce energy. The intermediates generated during this cycle are hydrophilic, acidic compounds, varying in acid strength from strong to moderately weak. Due to their low hydrophobicity, these compounds are not effectively retained on conventional reversed-phase columns such as C18 or C8. Using the Zodiac HST RP mixed-mode column, citric acid and related metabolites are successfully separated through a combination of reversed-phase and anion-exchange mechanisms. The method delivers strong retention, good peak shape, and robust separation of all hydrophilic acidic components. It is fully compatible with LC/MS detection and preparative chromatography, making it suitable for comprehensive analysis of TCA cycle intermediates and other polar acidic molecules.