An ultra rapid and sensitive high-performance liquid chromatographic method for determination of tissue and plasma...

An ultra rapid and sensitive HPLC method for measuring individual free amino acids in biological fluids has been developed by using o-phthaldialdehyde/3-mercaptopropionic acid as derivatization agent and employing 3-micron-particle-size reversed-phase columns. Resolution of the amino acid derivatives is accomplished with an acetonitrile gradient in 12.5 mM sodium phosphate buffer, pH 7.2. These conditions facilitate separation of the 23 major tissue free physiological amino acids in the lower picomole range in less than 13 min. Muscle, liver, and kidney free amino acid concentrations, as determined by HPLC, are in the expected physiological range and compare favorably with those obtained by conventional amino acid analyzer.