Abstract:Objective To establish a separation method for reference substance of mulberroside A from the root bark of Morus alba.Methods Ethanol-extract of the root bark of M.alba was isolated and purified by maeroporous resin,silica gel,and Sephdex LH-20 column chromatography.The purity of mul- berroside A was identified by TLC and ELSD-HPLC.Its structure was identified by spectral analysis.Re- sults The compound was completely separated from the root bark of M.alba.The purity of the reference substance was not less than 98.5%.Conclusion The compound prepared by the above-mentioned method is accorded with the relative demands of chemical reference substance in Chinese materia mediea.It can be used as a reference substance for the quality control and the research of herbal medicine.