Online hyphenation of centrifugal partition chromatography with countercurrent chromatography (CPC-CCC) and its application to the separation of saturated alkylresorcinols

Abstract

Centrifugal partition chromatography (CPC) and countercurrent chromatography (CCC) are two preparative techniques mainly used for the isolation and purification of natural products. While CPC benefits from a larger sample capacity, CCC typically provides better peak resolutions and hereby higher purities. In this study, we aimed to combine both advantages by the direct linking of CPC and CCC which was achieved by installation of switching valves and connection tube. The hyphenated CPC-CCC setup was tested with major alkylresorcinols which were obtained from a transesterified and hydrogenated rye extract. Injections of 1- and 5-g samples into the individual CCC system confirmed the limited sample capacity because of immediate flooding with the 5-g sample (total loss of stationary phase). In comparison, the CPC system was stable with 5- and 10-g samples but the peak resolution with 1-g sample was poorer than with the CCC system. Injections of 5- and 10-g samples into the CPC-CCC system were successful. However, a sample load of 10 g resulted in lower purities of the alkylresorcinols (80% or less) due to peak tailing. By contrast, injection of 5-g sample provided high amounts of ~ 1.2 g alkylresorcinols with purities of > 95%.