Separation and purification of peptides by high-speed counter-current chromatography

Preparative separations of peptides have been accomplished using high-speed counter-current chromatography. This has been made possible by the use of a particular solvent system that does not exhibit solvent carryover at high speed flow and centrifugation conditions. The solvent system composed of tert.-butyl methyl ether, n-butanol, acetonitrile and aqueous trifluoroacetic acid which can be adjusted in volume ratios and percent acid is employed in two instruments for counter-current chromatography. The cross-axis coil planet centrifuge was used for separation of various dipeptides. Superior resolution was obtained with the multi-layer coil planet centrifuge in the separation of six dipeptides and in the preparative purification of two synthetic peptides.