Abstract: We established a onestep preparation approach of high purity cis-and trans-2, 3, 5, 4′tetrahydroxystilbene-2-O-β-D-glycoside reference substance from Polygonum multiflorum Thunb.We initially extracted crude 2, 3, 5, 4′tetrahydroxystilbene-2-O-β-D-glycoside from Polygonum multiflorum Thunb.,then refined and purified it with preparative high performance liquid chromatography of acetonitrile andwater ratio of 25∶75 (V/V), and eventually acquired cis-and trans- 2, 3, 5, 4′tetrahydroxystilbene-2-O-β-D-glycoside. Their structures were identified by ESIMS,1Hand13CNMR. Their purities of 98.90% and 99.66%were determined with HPLC, which satisfiedthe requirement of reference substances. The method is simple, rapid, well reproductive, and has high extraction purity. It is therefore suitable for the preparation of cis and trans 2, 3, 5, 4′tetrahydroxystilbene2OβDglycoside reference substances.

Key words: 2, 3, 5, 4′tetrahydroxystilbene-2-O-β-D-glycoside, Polygonum multiflorum Thunb., preparation