Preliminary Study On The Processing Technology Of Bo-chrysanthemum And Its Chemical Composition And Liver Protective Effect Before And After Processing

ObjectiveChrysanthemum comes from the dried flower heads of Chrysanthemum morifolium Ramat.It has the functions of dispersing wind,clearing heat,calming liver,brightening eyes,clearing heat and detoxifying.Bo-Chrysanthemum is mainly produced in Bozhou,Anhui Province,which is one of the top ten medicine Anhui Province.Raw Chrysanthemum are often used in clinical,but it is slightly cold and easy to damage the spleen and stomach.After being fried,it can alleviate its coldness,which is beneficial to the use of the spleen and stomach discomfort.Mr.Xu Jingshi,a master of national medicine in the First Affiliated Hospital of Anhui University of traditional Chinese medicine,is good at using fried Chrysanthemum in clinical,which is mostly used in the prescriptions of yin deficiency of liver and kidney,deficiency of water and hyperactivity of fire,and unclear vision and material.The purpose of this study is to optimize the frying technology of Chrysanthemum,and to analyze the differences of chemical components and its protective effect on CCl₄induced acute liver injury in mice before and after frying,to provide a reference for clinical rational selection of Chrysanthemum processed products.Method1 Optimization of Chrysanthemum frying technology and comparison of“quality”and“quantity”before and after processingIn this experiment,multi-index orthogonal experiment method was used,and the temperature and time of frying were taken as the factors to investigate.The contents of chlorogenic acid,luteolin,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,luteolin,apigenin and total flavonoids in Chrysanthemum were used as the evaluation indexes,combined with the appearance of Chrysanthemum,the best frying process of Chrysanthemum was optimized.Based on the best technology,the fingerprints of raw and fried Chrysanthemum were established by UPLC.The cluster analysis and principal component analysis of raw and fried Chrysanthemum were conducted using SIMCA13.0.3 software.The content changes of 6 components before and after the Chrysanthemum frying were compared.2 Study on chemical composition difference of Chrysanthemum before and after fryingBased on the establishment of UPLC fingerprints of raw and fried Chrysanthemum and the quantitative analysis of six components before and after frying Chrysanthemum,ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry?UPLC-Q-TOF-MS?was used to analyze the chemical composition of Chrysanthemum before and after frying.Combining UNIFI software with Mass Lynx V4.1 software,and according to principal component analysis?PCA?,orthogonal partial least squares discriminant analysis?OPLS-DA?and variable importance projection?VIP?data statistical analysis methods to find components with significant differences before and after frying.3 Protective effects of Chrysanthemum before and after frying on CCl₄induced acute liver injury in mice90 male mice of Kunming were randomly divided into 9 groups,namely normal group,model group,biphenyl double fat group?positive control?,raw Chrysanthemum high,medium and low dose group,fried Chrysanthemum high,medium and low dose group,10 in each group.Each administration group was administrated intragastrically for 7 days.The normal group and the model group were administrated with the same amount of normal saline.1 hour after the last administration,except the normal group,each administration group was intraperitoneally injected with 0.2%CCl₄olive oil solution?0.1 m L/10 g?to establish a model of CCl₄induced acute liver injury in mice.After fasting for 24 hours,the eyeball of mice was removed to take blood,and the mice were dislocated and killed and the liver was separated.The contents of AST and ALT in serum,SOD and MDA in liver tissues were determined in combination with liver histopathological changes to study the protective effect of Chrysanthemum on CCl₄induced acute liver injury in mice before and after frying.Result1 Optimization of Chrysanthemum frying process and comparison of"quality"and"quantity"before and after fryingThe optimum frying technology of Chrysanthemum was the frying temperature of130?,frying time of 8 min,the color of fried Chrysanthemum was dark yellow.The fingerprints of raw and fried Chrysanthemum were established,15 common peaks were calibrated,the retention time difference of each chromatographic peak was small,but the relative peak area changed greatly before and after frying.Cluster analysis and principal component analysis obviously divided raw and fried Chrysanthemum into two categories,indicating that there are differences in internal quality before and after frying.The results showed that the content of 6 components of Chrysanthemum changed significantly before and after frying.After frying,the contents of chlorogenic acid,3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid decreased significantly,the contents of luteolin,luteolin and apigenin increased significantly.2 Study on chemical composition difference of Chrysanthemum before and after fryingA total of 28 chemical components were identified in raw and fried Chrysanthemum,and 18 different components were identified.Combined with the results of the first chapter,the content of 6 components of Chrysanthemum changed significantly before and after frying,and a total of 20 components with significant differences were analyzed.The content of 13 ingredients increased after frying,respectively,cynaroside,luteolin,apigenin,1,3-dicaffeoylquinic acid,apigenin-7-o-glucoside,diosmetin-7-o-rutinoside,acacetin-7-o-glucoside,acacetin-7-o-glucuronide,hesperetin,gallic acid,isorhamnetin,eriodictyol,scopoletin.The content of 7 components decreased after frying,respectively,chlorogenic acid,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,diosmetin-7-o-glucuronide,diosmetin-7-o-beta-d-glucopyranoside,luteolin-7-o-glucuronide,apigenin-7-o-glucuronide.3 Protective effects of Chrysanthemum before and after frying on CCl₄induced acute liver injury in miceCompared with the model group,the high,medium and low dose groups of raw and fried Chrysanthemum significantly reduced the contents of liver index,AST and ALT in serum,and increased the content of SOD.High and medium dose groups of raw and fried Chrysanthemum can significantly reduce the MDA content in liver tissue.The results of liver histopathology showed that the degree of liver tissue damage was improved in the high,medium and low dose groups of raw and fried Chrysanthemums,the hepatic cord gradually appeared,the infiltration of inflammatory cells decreased,and the swelling of the nucleus decreased.The results showed that both raw and fried Chrysanthemum could alleviate CCl₄induced acute liver injury in mice in clinic.ConclusionThe optimized Chrysanthemum frying technology was stable and feasible.The chemical compositions of Chrysanthemum before and after frying has the difference of"quality"and"quantity".Both raw and fried Chrysanthemum have protective effects on acute liver injury induced by CCl₄in mice.The results of this study will provide reference for the establishment of Chrysanthemum frying technology and the rational selection of Chrysanthemum processed products.