Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography.

This paper aims to be an aid to those chemists who are interested in utilizing high-speed counter-current chromatography (HSCCC), which is free of irreversible adsorption and offers high resolution comparable to column chromatography. It explains the selection of HSCCC conditions step by step including the selection of two-phase solvent systems, determination of partition coefficient (K) of analytes, preparation of two-phase solvent system and sample solution, selection of elution mode, flow rate, rotation speed, and on-line monitoring of the eluate. The paper covers both standard HSCCC and pH-zone-refining CCC techniques. Technical terms (italic) unfamiliar to the beginner are comprehensively explained in Glossary. Various examples of two-phase solvent systems used in HSCCC are listed in Appendices A and B. The commercial sources of HSCCC and other CCC instruments are described in detail in the study edited by Berthod [A. Berthod (Ed.), Counter-current Chromatography, Elsevier, Amsterdam, 2003].