| Menthae Haplocalycis herba,one kind of Chinese edible herbs,has been widely adopted for the clinical use in China for more than two thousand years.It has the efficacy of evacuating wind and heat,clearing the head,swallowing,penetrating the rash,dispersing the liver and so on.It is mainly used in the clinic of the wind and heat cold and the beginning of the wind and temperature.The traditional multi water decoction of mint,which is often followed,indicates that the volatile and non volatile components in the Menthae Haplocalycis herba water extract are both the material basis for its efficacy.However,the current research on the chemical composition of Menthae Haplocalycis herba mainly focuses on the volatile components and lacks the systematic characterization of the chemical components in the Menthae Haplocalycis herba aqueous extract.The process of metabolism in the Menthae Haplocalycis herba aqueous extract is not clear,which restricts the basic research of its pharmacophore.In order to preliminarily explore the material basis of Menthae Haplocalycis herba water extract,the methods based on ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap)and GC-MS were used to establish a rapid screening and identification method for the chemical constituents of Menthae Haplocalycis herba aqueous extract,and the chemical constituents and metabolism in the Menthae Haplocalycis herba aqueous extract were studied.The main contents and results are as follows:1 Rapid identification and identification of Chemical Constituents of aqueous extracts from Menthae Haplocalycis Herba by UHPLC-LTQ-Orbitrap and GC-MS methodsThe methods based on ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap)and GC-MS were developed for a comprehensive study of the chemical constituents in Menthae Haplocalycis herba aqueous extracts.Different types of compounds’ fragment patterns were speculated and summarized according to the relevant references and literatures.For known compounds,it were exactly identified by extract ion chromatograph and reference substances;for unknown compounds,it were rapidly identified based on accurate mass,fragmentation pattern,multistage mass fragments,and data reported previously.Based on the accurate mass measurement(<5 ppm),MS/MS fragmentation patterns,different chromatographic behaviors,an important parameter Clog P value and compared with the standards and references,85 compounds were unambiguously identified or tentatively characterized,including 38 flavonoids,16 phenolic acids,24 terpenoids and 7 phenylpropanoids.At the same time,GC-MS was used to identify the volatile oil of Menthae Haplocalycis Herba and water extract of Menthae Haplocalycis Herba.66 and 35 compounds were identified within 70 min of GC chromatographic retention time,respectively.The results systematically analyzed the chemical constituents in the water extract of Menthae Haplocalycis Herba,which laid a chemical foundation for further research on the metabolism of Menthae Haplocalycis Herba water extract.2 The metabolic study of main monomers from Menthae haplocalycis herba aqueous Extracts in ratsThe metabolism of the main monomers in Menthae Haplocalycis Herba aqueous extract,such as Rosmarinic acid,Diosmetin,Diosmin,Hesperetin and Hesperidin in rats were studied respectively after systematically understanding chemical composition of Menthae Haplocalycis Herba based on UHPLC-LTQ-Orbitrap high resolution liquid chromatography.The metabolic pathways of the monomer compounds were estimated by analyzing the fragmentation pattern of the monomers and identifying the metabolites of the monomeric compounds by combining the chemical components of Chinese medicine and the biotransformation process of metabolites.As a result,36,47,77,17 and 52 metabolites were identified after oral administration of Rosmarinic acid,Diosmetin,Diosmin,Hesperetin and Hesperidin,respectively.The main metabolic pathways of Rosmarinic acid in rats were first decomposed into small molecules,such as ferulic acid,trans caffeic acid,trans-m-coumaric acid,m-hydroxybenzoic acid and Danshensu,and then sulfation,glucuronic acid conjugation,glucose conjugation and so on;the main metabolic pathways of Diosmetin in rats were sulfation,glucose conjugation,glucuronic acid conjugation or the reactions of sulfation,glucose conjugation and glucuronic acid conjugation after demethylation/methylation;the main metabolic pathways of Diosmin in rats were decarbonylation,demethylation,demethoxyl,hydroxylation,hydroxylation and methylation,glucose conjugation,or dehydration,sulfation,glucuronic acid conjugation,and glucose conjugation on the basis of the hydrolysis of Diosmin;the main metabolic pathways of Hesperetin in rats are sulfation,glucuronic acid conjugation,glucose conjugation or sulfation after Hesperetin demethylation;the main metabolic pathways of Hesperidin in rats are dehydration,demethylation,demethoxyl,glucoside bonds or decarbonylation,sulfation,glucuronic acid conjugation,glucose conjugation on the basis of Hesperidin hydrolysis.The results not only enriched the metabolism of Rosmarinic acid,Diosmetin,Diosmin,Hesperetin and Hesperidin,but also laid a foundation for the identification of metabolites and metabolic pathways in the rats of Menthae Haplocalycis Herba water extract.3 The metabolic study of Menthae haplocalycis herba aqueous Extracts in ratsOn the basis of the metabolism of monomers in vivo,the metabolism of Menthae Haplocalycis Herba water extract in rats was studied by using UHPLC-LTQ-Orbitrap high resolution liquid chromatograph.The prototype constituents were discovered in rats’ biology samples by comparing blank sample,the retention time and MS fragments of components in vitro and vivo.By comparing the retention time and mass fractions of the components inside and outside the blank sample and the Menthae Haplocalycis Herba water extract and comparative analysis metabolites of monomers,the metabolites in the plasma,urine,and feces samples after oral administration of the Menthae Haplocalycis Herba water extract were rapidly selected,9,64 and 57 metabolites were detected from plasma,urine and feces samples were identified after oral administration of Menthae Haplocalycis Herba,respectively.The results enrich the metabolic connotation of Menthae Haplocalycis Herba water extract,and provide a metabolic basis for futher mechanism elucidation and futher in-depth metabolic research. |
