| Chicoric acid for active ingredients in natural medicine echinacea pHenolic acid compounds,with the deepening of Chicoric acid pHarmacology research,the role of more and more be known,related data show that Chicoric acid can through the intestinal absorption,but its absorption rate is very low how to more efficient to let the body to absorb,Chicoric acid triggered strong interest,the researchers through the study on the stability of Chicoric acid,understand to be absorbed in the intestine,but its mechanism is unclear,in this paper,the mechanism in the intestinal absorption were studied.The purpose of this study was to extract and purify Chicoric acid from echinacea root,to screen out the optimal extraction method,and to study the absorption mechanism of Chicoric acid in vitro.The main work of this paper can be divided into three parts,the main content and results are as follows.Firstly,the extraction technology of Chicoric acid was studied.Taking the yield of chicroic acid as the index,echinacea root powder(80 orders)as the raw material,ethanol as the extraction solvent for ultrasonic assisted extraction,the effects of solvent concentration,temperature,time,times and ratio of material to liquid on the yield were investigated.Through single factor experiment select test parameters,and then on the basis of single factor,choose solvent concentration(30%,40%,50%),extraction temperature(60℃,70℃,80℃),extraction time(1 h,2 h,3 h),and liquid ratio(1:12,1:15,1:18)four factors three levels orthogonal experiment design,in order to obtain the optimal extraction process.Secondly,the purification method of Chicoric acid was studied.The crude extract was further purified by dextran gel 1h-20 column chromatograpHic separation technology,and the chicroic acid monomer was obtained.Chicorianoic acid monomer was used as template molecule,4-vinyl pyridine as functional monomer,ethylene glycol dimethylenoic acid ester as crosslinking agent to prepare chicorianoic acid molecularly imprinted polymer,and this molecularly imprinted polymer was used as stationary pHase to enrich and purify echinacea root extract.Finally,the mechanism of absorption of Chicoric acid in vitro was studied.Caco-2 cells were cultured and cultured on Transwell plate to differentiate into dense and tightly connected monolayer cells.The caco-2 cell model was established,and the transmembrane resistance value of the model was measured to evaluate whether the model was successfully established.Acid in chicory again determined by MTT method is used to do cytotoxic test,by measuring each hole under 490 nm in the optical density(OD)value,the calculation cells,hence increment rate above 80%as no toxicity to cells,sure no cytotoxicity of chicoric acid concentration,and then choose different concentration(100 microns,200 microns,500 microns)chicoric acid Transwell experiment,selected 900 s,1800 s and 2700 s,3600 s and 5400 s,7200 s and 10800 s point in time,samples of the various points after UPLC detection,make medicine curve,P app value and outflow rate(Er)value were calculated to determine its absorption.Again using the same method of chitosan,glutathione and verapamil on cytotoxic test,analysis is safe and effective drug concentration,concentration of choice to join Transwell experiment was carried out,determining drug curve calculation P app value and Er,EDTA and chitosan have existing cellular pathways of transhipment,analysis of pH,verapamil and glutathione factors such as how to estimate the chicoric acid involved in transport and final analysis chicoric acid transport mechanism.The results showed that:1.The ratio of material to liquid was 1:18 with 50%ethanol at 70℃,and the extraction was assisted by ultrasound for 2h.The yield of Chicoric acid was 1.89 mg/g under the optimal conditions.2.After the extraction and separation of dextran gel LH-20,the content of Chicoric acid was determined by HPLC to 90.22%.After enrichment and purification of echinacea root powder by molecularly imprinted polymer,both Chicoric acid and caffeic acid with the same structure of parent nucleus could be enriched.3,Chicoric acid may be through with passive transport is given priority to,the transfer mechanism of active transport coordination,determine the EDTA and chitosan can improve the cumulative transfer traffic volume of different level,confirmed that passive transport transshipment of Chicoric acid participation,glutathione transferase and Na+channels participate in Chicoric acid transhipment,P-gp is not play a role in the transport,pH may also affect Chicoric acid transshipment. |
