Separation of a triterpenoid saponin mixture from Maesa lanceolata: semipreparative reversed-phase wide pore high performance liquid chromatography with temperature control

Abstract

A mixture of triterpenoid saponins derived from the dried leaves of Maesa lanceolata was separated, without structure deterioration, in its components. Seven fractions (I–VII) of high molecular weight (1234–1358) saponins were obtained on a semipreparative scale using wide pore reversed-phase high performance liquid chromatography with an acetonitrile–trifluoroacetic acid (500:0.3 w/w)–water–trifluoroacetic acid (391:0.3, w/w) gradient from 35 to 56% in 30 min. The mobile phase was cooled in an ice bath (0°C) during chromatography in order to prevent bubble formation and to improve the quality of the separation. Freeze-dried fractions IV, V, VI and VII were further separated using solvent systems developed for each of the fractions. Fourteen pure triterpenoid saponins were isolated in this way and their molar weight determined.