Abstract: In this study, Ilex rotunda Thunb sample was pulverized and extracted with 50% ethanol.Consequently,the crude product of syringin was obtained from the ethanol crude extract through purification with XDA-1 macroporous adsorption resin.In order to prepare syringin with the preparative HPLC,the initial chromatographic conditions of the preparative DAC-HB50 column (50 mm × 1 000 mm;the packing material:chromatorex C18,10 μm,300 g) were investigated with the analytical welchrom C18 chromatographic column (4.6 mm × 250 mm,10 μm) on the preparative HPLC.Furthermore,syringin was acquired from the crude product of syringin under the best loading amount that was sought through the optimization of the initial separation condition.Finally,the TLC and analytical HPLC analysis results showed that the final obtained product was syringin with the purity of 98.6%.The molecular structure was determined by single-crystal X-ray diffraction analysis.According to the data,the final obtained product was further confirmed as syringin.

Key words: preparative high performance liquid chromatography, syringin, Ilex rotunda Thunb, single crystal X-ray diffraction, purification