Analysis Of Ingredients From Evodia Rutaecarpa In Vivo Based On Hepatotoxicity

Chinese medicine is China’s cultural treasures.There is a long history of the use of traditional Chinese medicine for the treatment of various diseases.However,with the continuous development of modern technology and the introduction of Western medicine,the diversity of ingredients hindered Chinese medicine’s process of internationalization.Chinese medicine has complex diversity of ingredients which are still not clear of the principle of action.In order to implement the internationalization of Chinese medicine,we need to build them on the basis of its quality control standards desiderately.While in the process of the establishment of Chinese medicine quality control standards,the research of pharmacodynamic substance basis is indispensable which can help reveal the specific efficacy ingredients of traditional Chinese medicine.Traditional Chinese medicine Evodia rutaecarpa beginning in cold-induced febride,as in the article,belong to the traditional Chinese medicine WenLi medicine,in recent years,the study found that evodia rutaecarpa have liver toxicity effect.Our previous studies have shown that the serum ALT,AST and experimental liver body coefficient of the subjects increased very significantly(P<0.01),after ig administration with 50%ethanol extract of Evodia rutaecarpa(35g·kg-1·d-1)for consecutive 14d.Our investigation demonstrates that Evodia rutaecarpa had accumulation toxicity in liver.The hepatic injury caused by Evodia rutaecarpa of metabolism and substance basis are not yet clearly.This study investigated the stability of serum sample and urine sample on the different processing methods and storage time in rats with hepatic injury caused by Evodia rutaecarpa,using UPLC-QTof-MS,the analysis and the appraisal system of evodia rutaecarpa hepatotoxicity of active site in rat blood and urine after taking the drug metabolism of different parts of the ingredients into the blood components,thereby to reveal evodia rutaecarpa efficacy material base.Here are the main contents and results:In the first part,the drug-containing samples without pretreatment and that dried under nitrogen were stored at-20℃ for different time(0d,3d,5d,10d and 15d),and then detected by HPLC-DAD,respectively.All of the RSD values of common chromatographic peaks of serum samples dried under nitrogen were less than 20%,and that of untreated samples were less than 20%in 3d.The serum samples dried under nitrogen could be stored steadily at-20℃ for 15d.However,the untreated serum samples could be stored steadily at-20℃ for 3d.All of the RSD values of common chromatographic peaks of urine samples dried under nitrogen and that of untreated samples were less than 20%.The drug-containing urine samples without pretreatment and dried under nitrogen could be stored steadily at-20℃ for 15d.In the second part of the study,UPLC-QTof-MS was used to analysis the drug-contained serum of normal male rats after gavaged with the liquid of liver toxicityquasi of Evodia rutaecarpa.We marked 21 chromatogrphic peaks in the serum,and finally idetified 20 chemical constituents based on the MS information and parts of the standard substance information,including 15 prototype components:Dehydroevodiamine,Evodianinine,12aHydroxylimoni,Limonin,Dihydrorutaecarpine,Evodiamine,Rutaecarpine,1-Methyl-2-nonyl4(1H)-quinolone,1-Methyl-2-[(Z)-5-undecenyl]-4(1H)-quinolone,1-Methyl-2-[(4Z,7Z)-4,7tridecadienyl]-4(1H)-quinolone,1-Methyl-2-undecyl-4(1H)-quinolone,Evocarpine,1-Methyl2-[(6Z,9Z)-6,9-pentadecadienyl]-4(1H)-quinolone,Dihydroevocarpine,1-Methyl-2-[(Z)-9pentadecenyl]-4(1H)-quinolone;5 metabolites components:Hydroxy Dehydroevodiamine,NbDehydroevodiamine,14-Formyl-Dihydrorutaecarpine,Hydroxyrutaecarpine,Hydroxy evodiamine.In the third part,UPLC-QTof-MS was used to analysis the drug-contained urine of normal male rats after gavaged with the liquid of liver toxicityquasi of Evodia rutaecarpa.We mark 20 chromatogrphic peaks in the urine sample,and finally idetified 20 chemical constituents based on the MS information and parts of the standard substance information,including 15 prototype components:Synephrine,5-Methoxy-N,N-Dimethy ltryptamine,Dehydroe vodiamine,Evodianinine,12a-Hydroxylimoni,Limonin,6β-acetoxy-5-epilimonin,Evodiamine,Rutaecarpine,1-Methyl-2-nonyl-4(1 H)-quinolone,1-Methyl-2-[(Z)-5-undecenyl]-4(1H)quinolone,1-Methyl-2-undecyl-4(1H)-quinolone,Evodiaxinine,Dihydroevocarpine,1-Methyl2-[(Z)-9-pentadecenyl]-4(1 H)-quinolone,1-Methyl-2-pentadecenyl-4(1 H)-quinolone;5 metabolites components:Dehydroevodiamine glucuronide conjugation,Dehydroevodiaminesulfate,Hydroxy Dehydroevodiamine,Nb-Dehydroevodiamine,Hydroxyevodiamine.There were 14 same chemical constituents in drug-contained serum and drug-contained urine,including 11 prototype components:Dehydroevodiamine,Evodianinine,12aHydroxylimoni,Limonin,Evodiamine,Rutaecarpine,1-Methyl-2-nonyl-4(1H)-quinolone,1Methyl-2-[(Z)-5-undecenyl]-4(1 H)-quinolone,1-Methyl-2-undecyl-4(1 H)-quinolone,Evodiaxinine,Dihydroevocarpine,1-Methyl-2-[(Z)-9-pentadecenyl]-4(1 H)-quinolone;3 metabolites components:Hydroxy Dehydroevodiamine,Nb-Dehydroevodiamine,Hydroxyevodiamine.Based on above,we determined the hepatotoxicity effect active site of Evodia rutaecarpa,analyzed and identified the ingredients in the drug-contained serum,urine of rats with UPLCQTof-MS.There is important scientific value to guide the clinical use of Evodia rutaecarpa,at the same time,the results is of great significance to the medicinal material basic research.