In Vitro Metabolic Studies On The Isoflavones From Red Clover And Chickpea

Isoflavones were the main ingredients in Red Clover and Chickpea,which exhibited a wide spectrum of bioativites such as anti-tumor,anti-inflammatory,antioxidative and estrogenic effects.In this study,the isoflavones from Red Clover and Chickpea were investigated,including the following parts:(1)The extracting methods of isoflavones from Red Clover and Chickpea were established.Ononin,daidzein,irilone-4?-O-?-D-glucoside,sissotrin,pratensein,genistein,formononetin,irilone and biochanin A from Red Clover,and ononin,formononetin 7-O-?-D-glucoside 6''-O-malonate,biochanin A 7-O-?-D-glucoside 6''-O-malonate,formononetin and biochanin A from Chickpea were separated and purified with using macroporous resin and semi-preparative liquid chromatography,respectively.UPLC-PDA and UPLC-QTOF-MS were used to identify and testify the purity of the compound.The result showed that the purity of each compound was more than 95%.(2)UPLC-QTOF-MS was used to identify and elucidate the metabolites of isoflavones from Red Clover and Chickpea in the gastrointestinal model in vitro.It was found that isoflavones were stable in simulated gastric juice and intestinal juice.However,serveral metabolites were detected in human intestinal flora incubated samples.The results indicated that deglycosylation,hydrolysis,dehydroxylation,demethylation,hydroxylation,and methylation were the main metabolic processes of isoflavones in human intestinal flora in vitro.The metabolism of isoflavones from Red Clover and Chickpea were comprehensively studied and the in vitro metabolic pathways of isoflavones were elucidated.This study may provide the evidence for screening the active ingredients from isoflavones metabolites and provide the reference for in vitro gastrointestinal tract metabolism study of other natural drugs.