Preparation Of Acanthopanax Trifoliatus(L.)Merr.Ointment

Acanthopanax trifoliatus(L.)Merr.is a representative plant of Acanthopanax in Acanthopanax family,also known as Acanthopanax,Acanthus,etc.Acanthopanax trifoliatus(L.)Merr.is a common folk medicinal plant.Its root has the effect of dispelling wind and dehumidification,relaxing meridians and activating blood vessels.The leaves of Acanthopanax trifoliatus(L.)Merr.are used to treat carbuncle,sore,bronchitis,periarthritis of shoulder,etc.However,the basic research and development of Acanthopanax trifoliatus(L.)Merr.is far less than that of Panax ginseng,which belongs to Acanthopanax family.Taking Acanthopanax trifoliatus(L.)Merr.as the research object,this paper uses modern whitening theory to explore the extraction process,refining method and whitening mechanism of the active ingredients of Acanthopanax trifoliatus(L.)Merr.,to investigate its safety,and then to prepare a new dosage form of Acanthopanax trifoliatus(L.)Merr.ointment,which provides the basis and basis for further research of Acanthopanax trifoliatus(L.)Merr..The research content of this paper is as follows:1)Single factor test and response surface analysis were used to determine the best extraction process of Acanthopanax trifoliatus(L.)Merr..Based on the comprehensive score of tyrosinase activity inhibition rate and DPPH free radical scavenging rate,the water extract and alcohol extract of Acanthopanax trifoliatus(L.)Merr.from Anhui and Guangdong were compared.It was found that the tyrosinase inhibition rate and DPPH free radical scavenging rate of water extract and alcohol extract of Acanthopanax trifoliatus(L.)Merr.from Anhui were higher than those of Acanthopanax trifoliatus(L.)Merr.from Guangdong,that is,the comprehensive score of Acanthopanax trifoliatus(L.)Merr.from Anhui was higher.In addition,the comprehensive scores of water extract of Acanthopanax trifoliatus(L.)Merr.from two places were higher than that of alcohol extract.Then response surface analysis was used to determine the best extraction method of Acanthopanax trifoliatus(L.)Merr.:the extraction solution is water,the ratio of material to liquid is 1:11(g)/(m L),the extraction time is 2h,the extraction temperature is 75℃,the comprehensive score is 0.561.Through the verification experiment,the actual measurement value is 0.560,which is basically consistent with the prediction theoretical value of the model,which proves that the model is reliable.2)Through the adsorption test of macroporous resin,the comprehensive score of tyrosinase inhibition rate,DPPH free radical scavenging rate and dry extract yield rate was taken as the index to investigate the effect of HPD450,D101,AB-8 and D4020 macroporous resin static adsorption on the purification of Acanthopanax trifoliatus(L.)Merr.extract,so as to determine the best purification method of Acanthopanax trifoliatus(L.)Merr.extract.The results showed that the comprehensive score of the solution was higher than that of the solution.It is concluded that the macroporous resin adsorption method has a good effect in refining.Then,by comparing the comprehensive scores of the desorbed solution obtained by several macroporous adsorption resins,the best refining method of the extract of Acanthopanax trifoliatus(L.)Merr.was determined as follows:D4020 macroporous adsorption resin was used,and the proportion of drug solution:macroporous resin=10:3(m L:g)was used for refining,at this time,the impurities could be removed to the greatest extent and the best refining effect could be obtained.3)The safety of Acanthopanax trifoliatus(L.)Merr.refined solution was investigated by red blood cell hemolysis test and skin erosive resistance test in rats.According to the linear regression analysis of the hemolysis data of red blood cells,the linear regression equation of concentration X and hemolysis rate Y of the refined solution of Acanthopanax trifoliatus(L.)Merr.is Y=11.117X-0.0094,R~2=0.9536.When the concentration is 1g/m L,the hemolysis rate is 0.7980%<5%.The resistance value of the rat was 13.93kΩ,between 5 and 15kΩ.Then,the absorption of SRB was calculated according to the results of SRB staining,and compared with the positive control group,it was judged that the test substance of Acanthopanax trifoliatus(L.)Merr.refined solution had no skin corrosiveness and irritation.4)Through the inhibition of tyrosinase activity test,it was found that the effect of the refined solution of Acanthopanax trifoliatus(L.)Merr.on the inhibition of monophenol enzyme activity was not obvious,but on the inhibition of diphenolase activity was significant.The IC50 value of diphenolase inhibition was 61.351 mg/m L.In a certain concentration range,the inhibitory effect of tyrosinase activity was positively correlated with the concentration of Acanthopanax trifoliatus(L.)Merr.refined solution.The inhibition of the refined solution of Acanthopanax trifoliatus(L.)Merr.and L-dopa on the activity of diphenolase was reversible competitive inhibition.The inhibition constants KI and KIS of[EI]and[ESI]were calculated to be 0.0053mg/m L and 0.0107 mg/m L,respectively.KI<KIS,indicating that compared with the complex[ESI],the refined solution of Acanthopanax trifoliatus(L.)Merr.can bind the free enzyme[EI]more firmly and produce strong inhibition.5)Through response surface analysis,on the basis of preliminary matrix formula,taking stability,sensory performance and skin irritation as indicators,the best formulation of Acanthopanax trifoliatus(L.)Merr.was determined.In the formula,the emulsification system and thickening system have the greatest influence on the appearance,performance and stability of the ointment.The best emulsifier is cetearyl glucoside,the best thickener is sodium carboxymethylcellulose,the dosage range of Acanthopanax trifoliatus(L.)Merr.refined liquid is 1.0%～4.0%,and 2.0%has the closest p H value to human skin and the best sensory score.Then response surface analysis was used to determine the addition amount of cetearyl glucoside,sodium carboxymethylcellulose and Acanthopanax trifoliatus(L.)Merr.refining solution,which were 2.0%,1.0%and 2.0%,respectively.The results showed that the stability of the optimized formulation was evaluated by 90 days of centrifugation,cold and hot circulation,heat resistance and cold resistance.The results showed negative in the irritation test.It can be preliminarily considered that the Acanthopanax trifoliatus(L.)Merr.ointment with optimized formula will not produce skin adverse reactions to human skin.And through the human patch test,it is further verified that the Acanthopanax trifoliatus(L.)Merr.ointment has a low possibility of causing adverse reactions to human skin,which is in line with the scope of safe use.6)Through the study of quality control of Acanthopanax trifoliatus(L.)Merr.ointment,the quality of Acanthopanax trifoliatus(L.)Merr.ointment is qualified and effective.The linear regression equation of rutin was calculated by HPLC:Y=76.77X+34.673(R~2=0.9995)(n=5).There was a good linear relationship between rutin and peak area in the concentration range of0.5-2.5μg/m L.The average concentration of rutin was 1.532μg/m L,RSD was 1.15%.The linear regression equation of chlorogenic acid is Y=203.32X+106.99(R~2=0.9994)(n=5).The results show that there is a good linear relationship between chlorogenic acid and peak area in the concentration range of 2.0-10.0μg/m L.The average concentration of chlorogenic acid was5.558μg/m L,RSD was 2.24%.