Study On The Active Subsance Of Cyclocarya Paliurus Leaves And Schisandra Chinensis Fruits Based On Spectrum-effect Relationship

Traditional Chinese medicine（TCM）is rich in resources and has a long history of use.It has a good curative effect on many diseases,especially chronic diseases,and low side effects.Thus it has been the main source of lead compounds for new drugs discovery.The curative effect of TCM results from the synergistic effect of multi-components acting on multi-targets through multi-pathways.Revealing the material basis of the efficacy of a TCM is the premise of studying its quality control,pharmacological action and mechanism,which is of great significance to develop and utilize it.Spectrum-effect relationship study is to analyzing the relationship between the fingerprint of a TCM and its pharmacodynamic data through appropriate chemometric methods thus reflecting its internal quality and the corresponding efficacy.Spectrum-effect relationship study has been widely applied to controlling the quality TCM and revealing its material basis of the efficacy.In this dissertation,Cyclocarya paliurus（CP）leaves and Schisandra chinensis（SC）were taken as the research objects,and their active material bases were screened and identified based on spectrum-effect relationship study and liquid chromatography-mass spectrometry technique.The main research contents are as follows:（1）The study on spectrum-effect relationship of Cyclocarya paliurus leaves responsible forα-glucosidase inhibitory activity.After ultrasonic extraction with 60%ethanol,15 batches of CP from different areas were separated and purified by macroporous adsorption resin column,and the 70%ethanol elution fraction with the strongest inhibitory activity ofα-glucosidase was obtained.The chromatographic fingerprints of 70%ethanol elution fractions of 15 batches CP leaves were established by UPLC,and then 32 common peaks were obtained by similarity evaluation.Their IC₅₀values forα-glucosidase inhibition were measured by enzymatic reaction.Several multivariate statistical analysis methods including principal component analysis（PCA）,partial least square analysis（PLS）,grey relational analysis（GRA）were applied to analyze the correlation between the peak areas of common peaks and IC₅₀values,and then the spectrum-effect model was established.P10,P11,P18,P22 and P25 were assigned as the main chromatographic peaks contributing toα-glucosidase inhibitory activity.A total of 10 triterpenoids were identified from the 70%ethanol elution fraction of CP leaves by ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）analysis.This study reveals the effective components in CP leaves for inhibitingα-glucosidase,and provides a reference for its material basic research and development.（2）The study on spectrum-effect relationship of Schisandra chinensis fruits responsible for pancreatic lipase andα-glucosidase inhibitory activities.The UPLC fingerprints of 17 batches of SC fruits were established,and 14 common peaks were obtained by similarity analysis.The IC₅₀values for pancreatic lipase andα-glucosidase inhibitory activities were separately measured by enzymatic reactions.The spectrum-effect models were constructed by using multivariate statistical methods such as PCA,PLS and GRA to analysis the correlation between the peak areas of common peaks and IC₅₀values.The results showed that P1,P4,P6,P9,P10and P11 were the main chromatographic peaks contributing to pancreatic lipase inhibitory activity,and P1,P2,P4,P6,P9,P10,P11 and P13 were responsible for inhibitingα-glucosidase.Among them,P1,P4,P6,P9,P10 and P11 contributed greatly to inhibiting both pancreatic lipase andα-glucosidase.In comparison with the retention times and UV spectra of the standards,P1,P4,P9,P10,P12 and P14 were further identified as schisandrol A,schisandrol B,schisantherin A,schizandrin A,schizandrin B and schisandrin C,respectively.A total of 14 lignans were identified from the ethanol extract of SC fruits by UPLC-MS/MS.This study provides a certain experimental basis for elucidating the material basis of SC,and has important reference value for its development and utilization.