Separation of epimeric aromatic acid (-)-menthol esters by countercurrent chromatography using hydroxypropyl-β-cyclodextrin as an additive.

The separation of ten epimeric aromatic acid (-)-menthol esters by countercurrent chromatography with hydroxypropyl-β-cyclodextrin as the mobile phase additive was investigated, and methods for the analysis of all the epimeric esters by reversed-phase high-performance liquid chromatography were established. A biphasic solvent system composed of n-hexane/20-70% methanol containing 50 mmol/L of hydroxypropyl-β-cyclodextrin (1:1, v/v) was selected, which provided high separation factors for five of the epimeric esters, and successful separations by countercurrent chromatography were achieved. The complete separation of five pairs of epimeric ester was obtained with the purity being over 98% for each peak fractions, as determined by high-performance liquid chromatography. The recovery of each analyte from the eluted fractions reached around 80-88%.