A complete separation of dimethylaminoazobenzenesulphonyl-amino acids. Amino acid analysis with low nanogram amounts of polypeptide with dimethylaminoazobenzenesulphonyl chloride

Abstract

A chromatographical system has been developed to give a complete baseline separation of all dimethylaminoazobenzenesulphonyl (DABS)-amino acids for high-sensitivity amino acid analysis [Chang, Knecht & Braun (1981) Biochem. J. 199, 547--556]. The system, which uses a Merck RP-18 column with phosphate buffer (12 mM, pH 6.5)/acetonitrile mixture, allows reliable analysis of DABS-amino acids at the 1--2 pmol level. The accuracy of this new system is demonstrated by the composition analysis of two immunoglobulin light chains (214 amino acid residues) with differences at only three amino acid residue positions.