Comparative metabolism study on chlorogenic acid,cryptochlorogenic acid and neochlorogenic acid using UHPLC-Q-TOF MS coupled with network pharmacology

Chlorogenic acid(5-CQA),neochlorogenic acid(3-CQA),and cryptochlorogenic acid(4-CQA),usually simultaneously exist in many traditional Chinese medicines(TCMs).However,insufficient attentions have been paid to the comparative metabolism study on these three isomeric constituents with similar effects on anti-inflammation until now.In this study,a novel strategy was established to perform comparative analysis of their metabolic fates in rats and elucidate the pharmacological mechanism of antiinflammation.Firstly,diagnostic product ions(DPIs)deduced from the representative reference standards were adopted to rapidly screen and characterize the metabolites in rat plasma,urine and faeces using UHPLC-Q-TOF MS.Subsequently,Network pharmacology was utilized to elucidate their anti-inflammatory mechanism.Consequently,a total of 73 metabolites were detected and characterized,including 50,47 and 43 metabolites for 5-CQA,4-CQA and 3-CQA,orderly.Moreover,the network pharmacology study indicated that these three isomeric constituents and their major metabolites with similar in vivo metabolic pathways exerted anti-inflammatory effects through co-owned 20 biological processes,which involved 10 major signal pathways and 159 potential targets.Our study shed light on the similarities and differences of the metabolic profiling and anti-inflammatory activity among these three isomeric constituents and set an example for the further researches on the active mechanism of isomeric constituents existing in TCMs based on comparative metabolism study.