|Molecular Weight||134.088 g/mol|
Primesep 100 separates a mixture of dicarboxylic acids in ion-exclusion mode with a mobile phase of water, acetonitrile (MeCN, ACN), and sulfuric acid (H2SO4) with UV detection at 210 nm. Baseline resolution of fumaric, maleic, malic, and succinic acids is obtained in less than 8 minutes. The separation combines ion-exclusion and reversed-phase mechanisms in one method.
Primesep 200 retains and separates the organic diacids (malic, succinic, fumaric, and maleic) by a combination hydrophobic, reversed-phase interactions and ion exclusion. The separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.
Hydrophilic acids are separated on Obelisc N mixed-mode HILIC column. Seven carboxylic acids are separated based on their polarity and pKa values. Changes in ionization states of acids and stationary phase can be used to control elution order of organic and inorganic acids.
This is an alternative approach for the separation of the TCA cycle intermediates that was developed using novel hydrogen-bonding HPLC column. The organic nature of the mobile phase helps obtain a highly sensitive LC/MS compatible method.
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||Newcrom B, 4.6x150 mm, 5 µm, 100A|
|Mobile Phase||MeCN/H2O - 5/95%|
|Buffer||AmAc pH 5.0, Formic Acid|
|Flow Rate||1.0 ml/min|
|Detection||CAD (Corona) MS- compatible mobile phase|
|Class of Compounds||Acid, Hydrophilic, Ionizable|
|Analyzing Compounds||Glycolic acid, Succinic Acid, Methylmalonic acid, Malic Acid, Maleic Acid, Citric Acid, Tartaric Acid, Fumaric Acid, Malonic Acid|