Inexpensive, Computer-Automated HPLC for Ion Exchange Separation and Quantification of Amino Acids in Physiological Fluids

Abstract

An inexpensive, computer-automated HPLC for separation and quantification of amino acids in physiological fluids is described. The system offers fully automated equipment control, data collection, processing and storage capabilities. The component nature of the system and the software flexibility permit extensive system modification, accomodating a wide variety of different separatory procedures which are not possible with many dedicated amino acid analysers. The system uses a lithium-based ion exchange column with post-column o-phthalaldehyde derivatization. A time of 128 minutes, including column regeneration, is required for separation of all amino acids through to arginine. The advantages of post-column derivatization over pre-column derivatization methods for post-separatory amino acid techniques are discussed. Accurate quantification of radiolabel in amino acids is demonstrated.