摘要
Salvianolic acid A(Sal A),a water-soluble ingredient in Danshen,has various biological activities.Sal A and its impurities have similar physical and chemical properties,as well as strong reducibility;therefore,they are difficult to prepare and purify.In this study,high-purity Sal A was obtained by purification of sephadex chromatography and preparative chromatography.Furthermore,HPLC-DAD tandem ECD and HPLC-DAD tandem MS methods were used for non-volatile organic impurity analysis,ICP-MS method was used for non-volatile inorganic impurities and mass balance method and quantitative nuclear magnetic resonance were employed to certify the product.The structures of Sal A and its relative impurities were validated by nuclear magnetic resonance spectroscopy and mass spectrometry,and their contents were quantified as well.Following the principles of ISO Guides 34:2009 and 35:2005,a Sal A reference material was certified,covering homogeneity studies,stability studies,characterization,and uncertainty estimations.
基金
The authors acknowledge National Key R&D Program of China(Grant No.2016YFC1000900)
The Drug Innovation Major Project(Grant No.2018ZX09711001-001-015)should be substituted by Drug Innovation Major Project(Grant No.2018ZX09711001-003-022)
CAMS Innovation Fund for Medical Sciences(Grant No.2016-I2M-3-007)for financing this work.
