| Objective: In this paper,Houttuynia Cordata Thunb(H.Cordata)as a model,we analyzed the components related to the metabolism of 2-undecanone in H.Cordata by studying its total protein,water-soluble components and volatile oil components,and combined with supramolecular theory of traditional Chinese medicine,molecular connectivity index,dual modes of composition quality control and metabolism control,and we tried to establish a quality evaluation and control system suitable for the characteristics of traditional Chinese medicine,in order to realize the new idea and new pattern for comprehensive control of quality of traditional Chinese medicine.Methods: First,the total proteins in H.Cordata was extracted by Tris-HCL method,and the cure of standard protein was established by Braford method,and determination of protein contents in different parts of H.Cordata,the total proteins of H.Cordata was analyzed by SDS-PAGE electrophoresis,which laid the foundation for subsequent research on the metabolic pathway that related to 2-undecanone in H.Cordata.Then,the water soluble components and the volatile oil components of H.Cordata were extracted and analyzed.The metabolites of 2-undecanone in H.Cordata were investigated.The volatile oil from H.Cordata detected by GC/MS was taken as an index,calculation the molecular connectivity index of the volatile oil from H.Cordata.Finally,2-undecanone as the ligand,3-Glycidyloxypropyltrimethoxysilane as the silane coupling agent,the 2-undecanone affinity medium chromatography was prepared by using silica gel method,the GC,affinity medium and incubating method were used to capture the 2-undecanone metabolizing enzymes system in H.Cordata protein,so as to lay the foundation for further studies on the metabolism of other components in H.Cordata.Results: The curve equation of standard protein established by Braford method was Y=0.03535C+0.08445(R2=0.9995),the contents of protein in whole plant,leaves,roots and rhizomes of wild and fresh H.Cordata were 7.540 μg·m L-1,8.757 μg·m L-1,6.889 μg·m L-1 and 5.889 μg·m L-1 respectively,the contents of protein in rhizomes of H.Cordata is 4.522 μg·m L-1,which was bought form the market.The molecular weight of protein in H.Cordata was analyzed by SDS-PAGE electrophoresis,and the molecular weight range was 10-100 k Da.The fingerprint of water-soluble components in H.Cordata showed a lot of species,and the precision,repeatability and stability test were good,and the separation degree was also better.50 components of the volatile oil from H.Cordata were detected by GC/MS,the content of the top three were 2-undecanone,β-pinene and α-pinene,and calculated the molecular connectivity index of the volatile oil in H.Cordata.The MCI ranged from 2.480 to 12.822,the average MCI was 5.398,the MCI of 2-undecanone was 5.265,the MCI of houttuyfonate(decacyl acetaldehyde)was 5.731,the MCI of decanoyl acetic acid is 5.639.In the study of 2-undecanone metabolic node in H.Cordata,GC method showed that the composition group with a retention time of about 15 min were the components of 2-undecanone metabolism related ingredients.The 2-undecanone affinity medium was successfully prepared,and capturing the metabolic enzyme of 2-undecanone in H.Cordata,and its molecular weight was 10 k Da.Conclusion: The protein contents of wild and fresh H.Cordata is higher than the H.Cordata that brought from the vegetable market,and the contents of protein in wild and fresh H.Cordata is the whole plant,leaves,roots and rhizomes successively.The molecular weight of protein in H.Cordata is mostly large molecules.Houttuyfonate and decanoyl acetic acid were not detected in the water soluble components and the volatile oil components from H.Cordata,it was possible that houttuyfonate was oxidized to form the decanoyl acetic acid,which was further degraded to produce 2-undecanone,therefore,the content of 2-undecanone in H.Cordata’s volatile oil was the highest.The molecular weight of 2-undecanone metabolizing enzyme in H.Cordata was about 10 k Da,and then the metabolic pathway of 2-undecanone in H.Cordata is may be as follow: Decanoyl acetaldehyde 3-Oxododecanoic acid <=> 2-Undecanone + CO2,the metabolic enzyme is 3-oxolaurate decarboxylase. |
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