Retention correlation and orthogonality between reversed phase countercurrent chromatography and liquid chromatography based on solvent strength

Correlation of elution performance between reversed phase countercurrent chromatography and liquid chromatography was investigated using five selected natural components. Theoretical guidance for orthogonality of two-dimensional countercurrent chromatography and liquid chromatography was proposed. The difference in retention behavior between countercurrent chromatography and liquid chromatography was studied when the mobile phase was composed of methanol and water by measuring the partition behavior of five selected compounds in two typical biphasic solvent systems composed of n-hexane-ethyl acetate-methanol-water and chloroform-methanol-water. An orthogonal diagram between countercurrent chromatography and liquid chromatography was obtained by normalized treatment of the measured partition coefficients and capacity factors. The experimental results showed that each biphasic solvent system used for countercurrent chromatography had a high orthogonality with liquid chromatography when a specific volume ratio was used.