Preparative Isolation of Isorhamnetin from Stigma Maydis using High Speed Countercurrent Chromatography

Preparative high speed countercurrent chromatography (HSCCC) has been successfully used for the isolation and purification of isorhamnetin from Stigma maydis. This was achieved in two stages: the first separation was performed with a two phase solvent system composed of n-hexane-ethyl acetate-methanol-water (HEMW) at a volume ratio of 5: 5: 5: 5, yielding isorhamnetin at 65.6%, which is followed by the second run using a two phase solvent system composed of HEMW 5: 5: 6: 4, v/v. From 700mg of the crude extract, 11.8 mg of isorhamnetin was obtained at a high purity of 98%. The final identification was performed by MS, 1 H-NMR and 13 C-NMR spectra.