Ketoglutaric acid

Application description

This method focuses on the separation of compounds integral to the glutamate metabolomic cycle utilizing High-Performance Liquid Chromatography (HPLC) coupled with Mass Spectrometry (MS). The separation is conducted on an Amaze TH Tri-Modal Hydrophilic Interaction Liquid Chromatography (HILIC) Column, tailored to handle polar and ionizable compounds effectively. Compound Properties: Glutamate: As a neurotransmitter and key player in amino acid metabolism, glutamate exhibits water solubility and contains both acidic and basic functional groups, allowing its interaction under HILIC conditions. Alpha-Ketoglutarate (α-Ketoglutaric Acid): An intermediate in the TCA cycle, alpha-ketoglutarate is vital for cellular energy metabolism. It is water-soluble and possesses a keto acid functional group, contributing to its behavior in HILIC chromatography. Glutamine: A conditionally essential amino acid involved in protein synthesis and nitrogen transport, glutamine is water-soluble and contains both acidic and basic functional groups, suitable for separation under HILIC conditions. Glutamic Acid: Another neurotransmitter, glutamic acid is involved in various physiological functions. It is water-soluble and contains an acidic functional group, allowing for interaction in HILIC chromatography. Gamma-Aminobutyric Acid (GABA): Serving as a neurotransmitter that inhibits neuronal activity, GABA is water-soluble and possesses both acidic and basic functional groups, making it compatible with HILIC chromatography. LC-MS Conditions: Mobile Phase: Comprising a mixture of water and organic solvent, typically acetonitrile, supplemented with additives like ammonium formate or ammonium acetate to enhance ionization efficiency for MS detection. Gradient Elution: Employing a gradient of organic solvent to achieve optimal separation of compounds throughout the chromatographic run. Mass Spectrometry: Coupled with HPLC for sensitive and selective detection and quantification of metabolites based on their mass-to-charge ratio (m/z). By leveraging this method, efficient separation and detection of glutamate metabolomic cycle compounds can be achieved, facilitating comprehensive analysis of metabolic pathways and related biological processes.