Abstract
Microsequence analysis of polypeptides by the Edman procedure has become a standard and routine method for the protein chemist (1,2). This cyclic stepwise process is initiated by the reaction of phenylisothiocyanate (PITC) with the amino terminus of a polypeptide. Following this coupling step, the derivatized amino-terminal residue is selectively cleaved from the polypeptide and converted to a phenylthiohydantoin (PTH) amino acid. Various reverse-phase high performance liquid chromatography (HPLC) systems have been reported for the identification and quantitation of the resulting PTH amino acids (3–8). Recently, methods have been described which also use PITC derivatization for amino acid analysis (9,10). This procedure involves the reaction of PITC with amino acids to form phenylthiocarbamyl (PTC) derivatives and their subsequent identification and quantitation by reverse-phase HPLC.
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