Study On Processing Principle Of Liquor-processed Bufonis Venenum And Wine-broiled Zaocys Dhumnades

Objective: In the research progross of traditional Chinese medicine,liquor preparation method is widely used and important.At present,there are many researches on the liquor preparation method of Chinese medicine,it is generally believed that liquor preparation method can change the chemical composition of medicinal materials so as to change or ease the drug properties,enhance the efficacy of drugs,reduce drug toxicity and correct ugly odor.However,their mechanism still need to be further clarified.In this study,Bufonis Venenum and Zaocys dhumnades were selected as model drugs to explore the principle of reducing toxicity and deodorizing by processing with liquor and wine,in order to provide basis for theoretical and experimental research on the liquor preparation method of Chinese medicine.Method: 1.On the basis of the structure-toxicity relationship of 9 bufadienolides,the effects of temperature,light,solvent on chemical composition and toxicity were studied.The relationship between composition and toxicity was established to clarify the processing principle of liquor-processed Bufonis Venenum.1.1 With the chromatography and spectroscopic methods,the constituents of Bufonis Venenum were isolated and identified.Artemia toxicity experiments were used to explore the toxicities of bufadienolides from Bufonis Venenum1.2 HPLC method was used to analyze fingerprints and the contents of chemical constituents.Differential scanning calorimetry was used to determine the degree of protein denaturation of Bufonis Venenum with different processing methods.Acute eyes stimulation experiment and Artemia toxicities experiment were used to explore the toxicities of Bufonis Venenum with different processing methods.2.To compare volatile and non-volatile components between raw and processing Zaocys dhumnades,and explore the innate character of wine processing,to selecte technology of wine-broiled Zaocys dhumnades.2.1 Super Nose electronic nose technology and chemometrics were used to analyze the profile of volatile components in Zaocys dhumnades.The volatile components were identified by headspace gas chromatography-mass spectrometry(HS-GC-MS).LC-MS/MS and molecular network were used to compare non-volatile components in different samples.2.2 Orthogonal experiment was used to to select the best processing method of odor correction.Effects of different processing techniques on the color and odor of Zaocys dhumnades were investigated by Super Nose electronic nose technology and colorimetric method.Results: 1.19 compounds were obtained from Bufonis Venenum,including 17 bufogenins,1 bufoxin and 1 fatty acid.The toxicities of 9 bufadienolides were determined by acute toxicity test to Artemia.LC50 in 24 h of bufadienolides were2.54-74.48μg.m L-1.the number and position of hydroxyl groups,and the content of acetyl groups can affect their toxicities.2.The fingerprints of Bufonis Venenum with different processing methods were different.The contents of cinobufagin and resibufogenin from Bufonis Venenum after liquor-processing were lower than those of water-processing products.On the contrary,the content of cinbufotalin increased;Temperature and light have effects on the contents of the 3 compounds.The sample after freeze-dring with water showed a peak of heat absorption in the DSC spectrum and the strongest in eye irritation test of the rabbit model.The toxicities of Bufonis Venenum have been weakened by processing.The 24 h LC50 of Bufonis Venenum after liquor-processing was higher than that of water-processing products.3.The results of the electronic nose showed that the differences were significant between raw products and wine-broiled products,and the smell of wine-broiled products and wine were close.HS-GC-MS results showed that the relative content of aldehydes and 1-octene-3-ol were decreased after the stir-frying with wine,and those of heterocyclic and ester compounds were increased,and no sulfides were detected in wine-broiled Zaocys dhumnades.There were two common components in wine-broiled Zaocys dhumnades and wine,and the content of esters and alcohols are higher than raw and water-broiled Zaocys dhumnades.LC-MS/MS-molecular network showed that the species and contents of total non-volatile components in raw,water-broiled and wine-broiled products of Zaocys dhumnades were close.4.The results of ANOVA showed that the cooking time and temperature had no significant effect on the quality of wine-broiled products of Zaocys dhumnades.The optimal processing was determined to be 80℃ for the best frying temperature,2.5 h for the best frying time.The chromatic value showed that processing techniques could affect the color of wine-broiled Zaocys dhumnades,and the brightness value had a strong correlation with processing time.The results of the electronic nose showed that processing techniques had little effect on the odor composition and its contents.Conclusion: 1.Liquor-processing can denaturate protein.The constituents in Bufonis Venenum can change in temperature,solvent.the irritation degree,the contents of cinobufagin and resibufogenin of Bufonis Venenum were decreased after liquor-processing,and then the toxicity of Bufonis Venenum decreased.2.The contents of fishy materials,aldehydes 1-octene-3-ol and sulfides,were reduced by wine-broiling,but those of heterocycles and esters were increased significantly.It suggested that wine-broiling can correct ugly odor.The best technology of wine-broiled Zaocys dhumnades is to stir-fry 2.5 h under 80℃.