Separation of Compounds in TCA Cycle on Primesep D Column

The citric acid cycle (tricarboxylic acid cycle, Krebs cycle) is a key process in the metabolic pathway by which all aerobic organisms generate energy. Compounds generated during TCA are hydrophilic, acidic compounds. Some of the hydrophilic acids are very strong while others are relatively weaker. All compounds have very low hydrophobicity and do not retain by the reversed-phase mechanism on the C18 or C8 HPLC columns. Citric acid and related products were separated in reversed-phase, anion-exchange mode on the Primesep D mixed-mode column. All hydrophilic, acidic compounds are well separated and demonstrate good shape. This robust method can be used for analysis of components of the TCA cycle and other highly hydrophilic compounds. This method is fully compatible with LC/MS and prep chromatography.


Column Primesep D, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmFm, pH 3.0
Flow Rate 1.0 ml/min
Detection ELSD


Class of Compounds Acid, Hydrophilic, Ionizable
Analyzing Compounds Sodium, Succinic acid, Malic acid, Isocitric acid, Citric acid, Fumaric acid, Maleic acid
Application Analytes:
Citric Acid
Fumaric Acid
Isocitric Acid
Maleic Acid
Malic Acid
Succinic Acid