The application of fluorescein isothiocyanate and high-performance liquid chromatography for the microsequencing of proteins and peptides.

Amino acid derivatives of fluorescein isothiocyanate have been separated by reverse-phase high-performance liquid chromatography in 45 min, using a linear gradient formed from acetone and 10 mM sodium phosphate buffer (pH 7.0) at 60 degrees C. The fluorescence of the derivatives has been used for detection giving a sensitivity of less than 0.5 pmol for a single component. Applications of this method to the sequence analyses of egg white lysozyme and salmon beta-melanotropin are also described.