Study On Extraction Process Optimization And Activity Of Tanshinone From Salvia Miltiorrhiza

In this paper,ethanol extraction method and supercritical CO2 extraction method were used to optimize the extraction of tanshinone from Salvia miltiorrhiza.Combined with single factor and orthogonal experiments,the optimum conditions of ethanol reflux method were as follows: ratio of material to liquid 1:8,temperature 70°C,soaking time 1.0h,reflux time 2.0h,the highest total extraction rate of Salviae miltiorrhiza was 3.12%.The optimum conditions for the supercritical CO2 extraction method were as follows: pressure 35 Mpa,temperature 45°C,time 2.0h,entrainer flow rate 0.6mL/min,and the highest extraction rate of total ketones from Salvia miltiorrhiza.In contrast,the supercritical CO2 extraction method has higher extraction efficiency.Combined with liquid chromatography-mass spectrometry(LC-MS),the crude extr acts of Salvia miltiorrhiza extracted by the two methods were analyzed,and the fo ur main components in Salvia miltiorrhiza extract were identified as tanshinone IIA,tanshinone I,cryptotanshinone and dihydrotanshinone.According to the established four kinds of tanshinone standard curve,calculate the specific content of each comp onent.The results showed that the overall tanshinone extraction rate was tanshinone IIA> cryptotanshinone> tanshinone I> dihydrotanshinone.The AKTA purification system was used to further separate the crude Salvia milt iorrhiza extract.After separation,the components were identified by thin layer chro matography(TLC)and high performance liquid chromatography(HPLC).The result s showed that the four main tanshinone components were well separated.The purit y of the four tanshinones was determined by HPLC: tanshinone IIA : 96.685%,Tan shinone I: 93.083%,Cryptotanshinone: 94.968%,Dihydrotanshinone: 99.61%.The study of physicochemical activity of tanshinone specifically includes two asp ects: antibacterial activity and anticancer activity.In the antibacterial experiment,the results of CFU experiment showed that four tanshinones showed different degrees of bacteriostatic effect,in which cryptotanshinone had the strongest antibacterial abil ity.,followed by dihydrotanshinone and tanshinone I,tanshinone II A the weakest antibacterial effect.The MIC experiment further confirmed that cryptotanshinone has excellent antibacterial activity,in which the MIC value for Staphylococcus aureus was 20±0.08 ?g/mL,and the MIC value for Escherichia coli was 15±0.03 ?g/mL.Observation by scanning electron microscopy(SEM)confirmed that cryptotanshinone could destroy the cell membrane of E.coli and Staphylococcus aureus,causing ba cterial growth and reproductive dysfunction,and eventually leading to bacterial deat h.We used the active ingredient tanshinone IIA-the highest content of Salvia miltior rhiza,as an anti-cancer test subject.Because of its low water solubility,it cannot d irectly act on normal tumor cells.Therefore,a surfactant Poloxamer 188 was used t o encapsulate tanshinone IIA.The solid dispersion,after SEM characterization and s olubility test experiments,demonstrated that the water solubility of tanshinone IIA-P188-SD was 5 times that of tanshinone IIA.In the experiment of anti-cancer activity,human gastric cancer cells 7901 were tr eated with different concentrations of tanshinone IIA-P188-SD.After LIVE/DEAD e xperiments and flow cytometry,it was proved that tanshinone IIA-P188 has the effe ct of inhibiting the growth of cancer cells,and it is also resistant.The cancer activ ity increases as the drug concentration increases.