To establish a method for separating and purifying podophyllotoxin (PPT) from Sinopodophyllum hexandrum using microporous resin combined with preparative high-performance liquid chromatography (PHPLC).Fristly,static adsorption test was used for choosing the best type of microporous resin.Secondly,the optimum conditions (the load solution concentration,volume and flow rate of loading solution,and eluent concentration,eluent volume and eluent flow rate) were determined by dynamic adsorption experiment.Finally,PHPLC was used for isolating and purifying PPT.The confirmation of chemical structures was performed by ¹H NMR and ¹³C NMR.The optimum purification conditions of microporous resin was obtained as follows:the 60 milliliter of 0.46 mg/mL PPT extract was loaded on the resin column at flow rate of 3 mL/min and eluted with 120 milliliter of 80% ethanol at flow rate of 3 mL/min.PPT with high purity was obtained by preparative HPLC with a C₁₈ column (Megres)by using 34% acetonitrile in ultrapure water at a constant flow rate of 14 mL/min and a sample volume of 1mL.HPLC analysis showed that the purities of PPT > 98%,and the structure was confirmed as PPT.The method is simple,economic,efficient and can be applied to the separation and preparation of PPT from Sinopodophyllum hexandrum.

microporous resin;preparative high-performance liquid chromatography;isolation and purification;Sinopodophyllum hexandrum, podophyllotoxin