Separation of two quaternary protoberberine alkaloids in corydalis saxicola bunting by high-speed counter-current chromatography combined with silica gel column chromatography

A method was developed for the separation of two quaternary protoberberine alkaloids.After partitioned by different organic solvents,the water residue of the ethanolic extract of Corydalis saxicola Bunting was fractioned by silica gel column chromatography.The fraction contained dehydrocheilanthifoline and dehydrocheilanthifoline was chosen for the further separation by high-speed counter-current chromatography with two-phase solvent system composed of n-hexane-ethylacetate-n-butanol-ethanol-water-ammonia(0.4∶1.5∶4∶0.4∶5∶0.11 in volume ratio).The lower phase was used as stationary phase and the upper phase as mobile phase.As the result,9.28 mg of dehydrocheilanthifoline(purity 97.0%,HPLC) and 4.38 mg of dehydroisoapocavidine(purity 90.7%,HPLC) were obtained from 64.81 mg of sample.The structures of the isolated compounds were confirmed by the comparison of their retention time in HPLC and ESI-MS spectra with those of standard references.