Separation of peptides by spiral countercurrent chromatography

Spiral countercurrent chromatography (CCC) utilizes a new separation rotor composed of 8 high density polyethylene plates with spiral flow channels, mounted in the type-J coil planet centrifuge (CPC). Synthetic peptides were evaluated for partition coefficients in various solvent systems. Approximately 30mg amounts were chromatographed and the fractions analyzed for peptide content and purity. The solvent systems which contained sec-butanol or n-butanol and the mobile phase selected produced good purification. The stationary phase was highly retained over 60%, thus the spiral separation coil is able to retain these more viscous, low interfacial tension solvents that have not been able to be used well in the multi-layer coil for high speed CCC.