pH-Zone-refining centrifugal partition chromatography for preparative isolation and purification of steroidal glycoalkaloids from Solanum xanthocarpum

摘要

pH-Zone-refining centrifugal-partition chromatography (CPC) was successfully applied in the separation of complex polar steroidal glycoalkaloids of close Rf values, directly from a crude extract of Solanum xanthocarpum. The experiment was performed with a two phase solvent system composed of ethyl acetate/butanol/water (1:4:5 by volume) where triethylamine (5 mM) was added to the upper organic mobile phase as an eluter and TFA (10 mM) to the aqueous stationary phase as a retainer. Separation of 1 g of crude extract over CPC resulted in two distinct pH-zones. The fractions collected in pH-zone i afforded 72 mg of solasonine while the fractions collected in pH-zone ii were slightly impure, hence were purified over medium pressure LC, which afforded 30 mg of solasonine and further 15 mg of solamargine (SM). The steroidal glycoalkaloids, SM and solasonine were isolated in 93.3 and 91.6% purity, respectively. The isolated alkaloids were characterized on the basis of their H-1, C-13-NMR, and ESI-MS data.