Isolation and purification of acteoside and isoacteoside from Plantago psyllium L. by high-speed counter-current chromatography

Two isomeric phenylethanoid glycosides, acteoside and isoacteoside were isolated and purified from the seeds of Plantago psyllium L. for the first time by high-speed counter-current chromatography (HSCCC) using a solvent system consisting of ethyl acetate–water (1:1, v:v). By injecting 200 mg of the n-butanol extract of P. psyllium for five consecutive times, the two-step HSCCC procedure yielded a total of 165 mg of acteoside and 17.5 mg of isoacteoside from 978 mg extract. The recovery rates for acteoside and isoacteoside were 90 and 84%, respectively, and the purities were 98 and 94%, respectively. The HSCCC fractions were analyzed by HPLC and the structures were identified by UV, LC–APCI–MS in negative ion mode, and confirmed by NMR experiments.