ABSTRACT
Objective To establish a method for preparing the reference substances of hyperin and isorhamnetin- 3- O- galactoside. Methods Methanol- extract of Fructus Evodia was isolated and purified by combining solvent extraction with column chromatography and recrystallization. Hyperin and isorhamnetin- 3- O- galactoside were identified by mass spectrometry (MS), 1H nuclear magnetic resonance (1HNMR ) and 13C nuclear magnetic resonance (13CNMR) and their content was determined by HPLC. Results The purity was 99.6 % and 98.0 % for hyperin and isorhamnetin- 3- O- galactoside respectively. Conclusion This method is simple cancl effective to yield high- purity products. And these high- purity products can be used as reference substances for the research of herbal medicine.