Chromatographic separation of n-butyl N-trifluoroacetyl esters of amino acids

Abstract

A number of commercially available polar stationary phases were investigated to find suitable column substrates that would completely separate the n-butyl N-trifluoroacetyl ester derivatives of the 20 natural protein amino acids. Columns of two mixed stationary phases were investigated, as well as columns packed with two liquid phases in series. Information is also given on different column lengths and concentrations of two stationary phases and use of temperature programing. Complete resolution of a 20-component mixture of the n-butyl N-trifluoroacetyl ester derivatives was obtained with a mixed stationary phase column of 0.75 0.25 w w % of DEGS EGSS-X . The method has been applied to hydrolyzates of proteins and good chromatograms were obtained for bovine serum albumin and κ-casein.