Accumulation And Transformation Of Chemical Constituents In Salvia Miltiorrhiza

Objective:(1)To investigate the dynamic changes of the chemical constituents in different drying processes of Salvia miltiorrhiza and the leaves from S.miltiorrhiza,reveal the accumulation and transformation of the chemical constituents in them,and provide some references for the production processing and drying methods of Salviae Miltiorrhizae Radix et Rhizoma and Salviae Miltiorrhizae Folium.(2)To investigate the differences and transformation of free and bound phenolic acids before and after the drying process of S.miltiorrhiza,further elucidate the accumulation and transformation of phenolic acids during the drying process of fresh S.miltiorrhiza,and reveal that salvianolic acids are the products induced by drying stress after harvesting of fresh S.miltiorrhiza.Methods:(1)The content of main components was used as the index in this study to evaluate the processing method,drying method,correlation between dehydration rate and component content for fresh S.miltiorrhiza.In addition,the sealed and unsealed parallel control groups were set to carry out verification test during the drying process.UPLC method was used for determination of seven main components including rosmarinic acid,lithosperic acid,salvianolic acid B,cryptotanshinone,tanshinone?,methylene salianolate and tanshinone?_Ain S.miltiorrhiza.(2)The content of main components was used as the index in this study to evaluate the processing method,drying method,correlation between dehydration rate and component content for the leaves from fresh S.miltiorrhiza.In addition,the sealed and unsealed parallel control groups were set to carry out verification test during the drying process.UPLC method was used for determination of four main components including Rutin,isoquercitrin,rosmarinic acid and salvianolic acid B in the leaves from S.miltiorrhiza.(3)we studied hydrolysis method,hydrolysates and hydrolysis regularity of phenolic acids in S.miltiorrhiza.UPLC method was used to determine four main hydrolysates of bound phenolic acids,namely danshensu,SMND-309(caffeic acid dimer),caffeic acid,przewalskinic acid A(prolithosperic acid),and three main free phenolic acids in S.miltiorrhiza,namely rosmarinic acid,lithospermic acid,salvianolic acid B.Results:(1)The results showed that the fresh S.miltiorrhiza contained low levels of phenolic acid,and the content of phenolic acid increased significantly with the increase of dehydration rate during drying process,while the change of tanshinone was not obvious.In the comparison of three drying methods,we found that drying at 50?was better than drying in the sun,and drying in the sun was superior to drying in the shade.(2)The content of the phenolic acids and the flavonoids were present in very low concentrations in the leaves from fresh S.miltiorrhiza.However,during the drying process of different drying methods,the contents of each component increased significantly with an increase of dehydration rate.The assay results of the three different drying methods demonstrated that the drying in the shade is better than drying in the sun,and drying in the sun is superior than drying at 50?.(3)The results of the acid-base hydrolysis experiment of salvianolic acid showed that the alkaline hydrolysis effect was significantly better than acid hydrolysis.The optimal alkaline hydrolysis condition was hydrolysis at 70°C for 4 h with 2mol·L~-11 NaOH solution containing 1%ascorbic acid(Vit C).The hydrolysates of free phenolic acids were the same with the hydrolysates of bound phenolic acids.Fresh S.miltiorrhiza contains a low level of free phenolic acids and a high level of bound phenolic acids,which were exactly opposite to dried S.miltiorrhiza.Conclusion:(1)The content of phenolic acids and flavonoids in fresh S.miltiorrhiza and the leaves from S.miltiorrhiza leaves was very low,which was formed in a large amount during the process of drying stress after harvest,indicating that salvianolic acid and flavone were the products induced by drying stress after harvest.(2)Drying at 50?was the best drying method for S.miltiorrhiza and drying in the shade was the best processing method for the fresh leaves from S.miltiorrhiza.(3)It was suggested that a large amount of bound phenolic acids was accumulated during the growth of S.miltiorrhiza.These bound phenolic acids were coupled with polysaccharides on the cytoderm through ester bonds to form insoluble phenolic acids,which was not easy to be detected by conventional methods.However,during drying and dehydration processes,the bound phenolic acids were converted to a large amount of free phenolic acids under the action of the relevant enzyme.