Preparation Of Paeoniflorin Alcohol Body And Evaluation Of Its Transdermal Performance

Arthritis is called"arthralgia syndrome"in Chinese medicine,which is one of the main threats to the aging society diseases.Arthritis has a high incidence of longer duration,protraction and clinical drug dependence big features.Paeoniflorin has a significant effect in the treatment of rheumatoid arthritis and other aspects.But its poor fat-soluble make itself cannot directly through the biofilm,which results in low bioavailability and Influnces the efficacy on the play Chinese medicine usually use the traditional skin topical administration in the treatment of arthritis,which limits the drug through the skin by the penetration into the stratum corneum skin deep.Ethosomes as a highly permeable carrier can promote transdermal absorption of paeoniflorin external preparation.Paeoniflorin(paeoniflorin,PF)is from peony plants and is clinically used to treat rheumatoid arthritis as a main active ingredient of a total glucosides of paeony(total glucosides ofpaeony,TGP).However,its oral bioavailability of paeoniflorin is low and its penetration rate of conventional topical administration of drugs is also very low.Therefore,this study is to develop a kind of hypertonic administration carrier "Ethosomes"The study compared and screened the ethanol injection method,a film dispersion method and reverse evaporation method with index of morphology,stability,particle size distribution and the entrapment efficiencies,and finally prepared paeoniflorin ethosomes by reverse phase evaporation.Firstly experiments performed single factor,then optimize the preparation process of paeoniflorin Ethosomes by Box-Behnken response surface method with the index of the entrapment efficiencies.Experiments combined the optimized quality ethosomes with gels.Finally this study compared the capacity of permeation in vitro percutaneous absorption and transdermal administration absorption of paeoniflorin aqueous gels and paeoniflorin ethosomes gels,then to evaluate and explore the feasibility and necessity of constructing paeoniflorin ethosomesThe main experimental results are as follows:1.The establishment for the determination method of drug in paeoniflorin ethosomesThe study established the determination method of drug in paeoniflorin ethosomes with the idex of paeoniflorin The method had a good resolution,better stability and reproducibility.2.The establishment of the determination method of entrapment efficiency methodBy comparing the advantages and disadvantages of separation method,such as dextran gel column chromatography separation method and low speed-ultrafiltration centrifugal separation method,the study selected the low-speed centrifugation-ultrafiltration as the measuring method for determine the entrapment efficiency of paeoniflorin ethosomes,which was simple and its parallel operation was also good.In low-speed centrifugation-ultrafiltration method,experiment investigated the effect of time on the UF paeoniflorin transmittance,and finally the ultrafiltration time was set to 30min;By examining the impact of drugs on lipid concentration by ultrafiltration,experiments got the result which was that paeoniflorin has no adsorption on blank ethosomes;The average particle diameter of the ultrafiltration liquid was 0,which indicated that the use of ultrafiltration membrane ultrafiltration tube could completely trapped paeoniflorin ethosomes.Therefore,the choice of experimental low-speed centrifugation-ultrafiltration could be used to determine encapsulation efficiency of paeoniflorin ethosomes.3.Selectionof the preparation method of peony glycoside ethosomesFirstly phospholipid species were screened.When soybean phosphatidylcholine was S-100,the paeoniflorin ethosomes prepared by ethanol injection method,a film dispersion method and reverse phase evaporation method had a good preliminary stability Based on these study results,the experiment fixed the index of concentration of phospholipid,mass ratio of phospholipid to cholesterol,mass ratio of phospholipids to paeoniflorin,and compares and screens the ethanol injection method,a film dispersion method and reverse evaporation method with index of the entrapment efficiencies,finally prepared paeoniflorin ethosomes by reverse phase evaporation which resulted in high entrapment efficiencies4.Optimized formulation with Box-Behnken methodFactors affecting the reverse phase evaporation method included organic solvent type,the volume proportion of oil phase and water phase and the percentage of athanol.The single factor experiment tested the ratio of phospholipid to cholesterol,soy phosphatidylcholine concentration and concentration of the drug.When the phospholipid to cholesterol ratio of was 5:1,soybean phosphatidylcholine concentration was 13.6mg/mL,drug concentration was 2.8mg/mL,the maximum encapsulation efficiency was(36.87 ± 5.11)%.On the basis of the results of the single factor,phospholiid to cholesterol ratio(X1),phospholipid concentration(X2),paeoniflorin concentration(X3)were selected as factors,encapsulation efficiency was selected as the response.Experiment usesd Design-Expert 8.0 to get multivariate binomial regression;R2 obtained by fitting equation was 0.9777;ANOVA results showed that the case can fit the regression model very well and the experimental error was small,thus the model can be better used to accurately analyze and forecast the encapsulation efficiency.Optimal prescription levels of the value of each factor were phospholipid to cholesterol ratio(mg/mg)=5.03,phospholipid concentration(mg/mL)=17.62 mg/mL,paeoniflorin concentration(mg/mL)=5.19 mg/mL.Taking into account the practical feasibility of the experiment,the study set the level of the value of each factor which were the ratio of phospholipid to cholesterol(mg/mg)=5.0,phospholipid concentration(mg/mL)=18.0mg/mL,paeoniflorin concentratio(mg/mL)=5.2mg/mL.Test value of encapsulation rate of paeoniflorin ethosomes was(44.27 ± 0.27)%,which had an absolute deviation of 2.46%to the predicted value of 43.18%that was less than 10%,so the regression model can predict the analysis prescription of paeoniflorin ethosomes;The average particle size of the optimal prescription ethosomes was(212.1 ± 14.98)nm,polydispersity:index PDI was 0.16±0.02.5.The in vitro permeation study of paeoniflorin ethosomes and transdermal drug delivery researchIn vitro release results showed that release processes of paeoniflorin aqueous gels wasbetter than paeoniflorin ethosomes gel.Cumulative percentage within 14h of paeoniflorin inpaeoniflorin aqueous gels was(51.41±6.93)%,while in paeoniflorin ethosome gels was(49,73± 1.79)%.But in vitro release profiles,the release of paeoniflorin aqueous gel was fester and better than paeoniflorin ethosomes gel.6.Percutaneous penetrationPercutaneous penetration results of paeoniflorin ethosomes gel showed that the cumulative permeation amount was(361.67±69.98)?g·cm-2 and cumulative transmittance in 14h was(19.40±3.75)%;Percutaneous penetration results of paeoniflorin aqueous gel showed that the cumulative permeation amount was(205.28± 37.88)?g·cm-2 and cumulative transmittance in 14h was(9.72±1.80)%.The cumulative permeation amount and cumulative transmittance in 14h of paeoniflorin ethosomes gel were 1.76 times and 2.00 times of paeoniflorin aqueous gel,indicating that transdermal performance of paeoniflorin ethosomes gel was better than paeonflorin aqueous gel.