| Traditional Chinese medicine(TCM)has a long history in China,and it was mainly taken by oral administration.After oral administration,the multicomponents of TCM must go through gastrointestinal tract before being absorbed into hematological system.As we all know,the combination of drugs and targets is the mechanism of exerting the efficacy of the drug.And the drug molecules will be expelled from the body by metabolism.However,because Chinese medicine is mainly derived from animals and plants,most Chinese medicine has the characteristics of biological diversity,variability,complexity and so on.Besides,the interaction between absorption,metabolism,distribution,excretion,and components makes the composition of compounds more complex.And due to the toxic and side effects of the drug,the general drug concentration and the amount of metabolites in the body are very low.All of the factors have increased the difficulty of the study of Chinese medicine and also become an important barrier to the internationalization of Chinese medicine.Fortunately,with the development of modern science and technology,the analysis and detection technology has been continuously improved.The high sensitivity,accuracy and selectivity characteristics of ultra-high-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry(UHPLC-LTQ-Orbitrap)make it possible to analysis the biological samples.Therefore,this study taken the water extract of areca catechu(WEAC)and simotang oral liquid(SMT)as the research object,and analysis of their chemical constituents and metabolites by LC-MS/MS.Besides,we study the metabolic behavior of arecoline with different chemical constituents by using in vitro enzyme incubation,and the result may provide a useful methods for the study of compatibility of TCM with detoxification.1.Identification of the main constituents of SMT and WEAC.In the present study,a sensitive and efficient method based on UHPLC-LTQ-Orbitrap was developed to characterize the chemical constituents of SMT.According to the mass spectrometry data,chromatographic behaviors,reference standards and previous literatures,a total of 71 compounds were unambiguously or tentatively characterized,mainly including 26 flavonoids,14 alkaloids,4 coumarins,5 amino acids,7 organic acids,13 terpenoids and.2 other compounds.2.Screening and identification of the metabolites in rat plasma and urine after oral administration of WEAC.In this study,an UHPLC-LTQ-Orbitrap method was used to analyze the major components and their metabolites in rat plasma and urine after oral administration of WEAC.A total of 41 constituents had been identified,including 9 prototypes and 32 metabolites were identified in rat plasma and urine samples.And the chromatographic separations were performed on a Thermo Scientific BDS Hypersil C18 column 2.1 x 150 mm2,with 2.4-?m particle size.The mobile phases A and B comprised 0.1%formic acid in water and acetonitrile,respectively.3.Screening and identification of the metabolites in rat plasma and urine after oral administration of SMT.In this study,an UHPLC-LTQ-Orbitrap method was used to analyze the major components and their metabolites in rat plasma and urine after oral administration of SMT.A total of 38 constituents had been identified,including 12 prototypes and 26 metabolites were identified in rat plasma and urine samples.And the chromatographic separations were performed on a Thermo Scientific BDS Hypersil C18 column 2.1 × 150 mm2,with 2.4-?m particle size.The mobile phases A and B comprised 0.1%formic acid in water and acetonitrile,respectively.4.The in vivo process analysis of SMT.In this study,the method of in vivo and closed intestinal ring method combined with UHPLC-LTQ-Orbitrap technology was used to analyze the absorption and metabolism of SMT.A total of 18 constituents had been identified.And 15 components of the SMT were absorbed into the blood by the prototype,and 3 components were metabolized by intestinal enzymes or liver.5.Study on the attenuated effect of arecoline under different compatibility.To observe the metabolic status of arecoline in WEAC,SMT and in rat liver microsomes alone.The metabolic status of arecoline was studied by using in vitro incubation method in rat liver microsomes,either alone or in combination.The remaining concentrations of arecoline in the liver microsomal incubation solution were analyzed by high performance liquid chromatography to study their metabolic status.The results showed that there were significant differences in the metabolism of arecoline with different combination.Conclusions:The study of the results in this paper had identified the constituents of WEAC and SMT,also prediected the possible metabolism proess of the main compounds in WEAC and SMT,which had provided essential data for further safety assessment,pharmacological studies,and quality control studies of areca catechu and SMT.Meanwhile,the metabolism experiment of enzyme in vitro also provided valuable data for the study of the mechanism of drug compatibility of Chinese medicine. |
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