| Buddleja lindleyana Fortune belongs to the genus Buddleja lindleyana of loganaceae.It is a kind of folk Chinese herbal medicine,which is widely used in the treatment of asthma,cough,activating blood circulation,activating collaterals,removing blood stasis,relieving pain,and expelling insects in the body.At the same time,it is used as the traditional ethnic medicine of Tujia nationality to treat burns,tinea corporis,tinea pyonensis,itching skin,cold and wind.Taking Buddleja as the research object,the extraction method,purification method and whitening mechanism of Buddleja whitening active substances were explored by using modern whitening theory,and their safety was evaluated.Then Buddleja cream was prepared,which provided reference for the follow-up research promotion and scientific research industrialization of Buddleja and Chinese medicine cream.The experimental content and plan of this researchSingle factor and response surface methodology were used to optimize the extraction process of whitening active substances from Buddleja.Taking the comprehensive scores of tyrosinase activity test in vitro and antioxidant DPPH test in vitro as the evaluation indexes,the water extract and alcohol extract of Buddleja from Anhui and Jiangsu were compared.It was found that the comprehensive scores of water extract and alcohol extract of Buddleja from Anhui were higher than those of Buddleja from Jiangsu,and the comprehensive scores of water extract of Buddleja from Anhui and Jiangsu were higher than those of alcohol extract.The results of single factor test were analyzed by response surface analysis showed that water was the best extraction solvent,The best material-to-liquid ratio,extraction time and extraction temperature are 1:19(g/mL),84 min,41℃,respectively.At this time,the theoretical comprehensive score was 56.990.Through the verification experiment,The actual comprehensive score is 56.897,and there is almost no difference between the actual and predicted values,which verifies the accuracy and reliability of the model.Based on the good adsorption performance macroporous resin,taking the tyrosinase activity test in vitro,DPPH test in vitro and the comprehensive score of dry extract yield as indexes,the static adsorption effect of HPD450,D101,NKA and AB-8 macroporous adsorption resins was investigated to determine the optimal purification method.Compared with the comprehensive score of macroporous resin washing solution and desorbing solution,it was found that the comprehensive score of desorbing solution was higher than that of washing solution.Finally,it is determined that the best purification method of Buddleja extract is D101 macroporous adsorption resin was used,and the ratio of liquid:macroporous resin=10:3(mL:g)was used for refining.At this time,the impurities were removed to the greatest extent,the functional components were retained,and the best refining effect was obtained.The safety of refined Buddleja solution was investigated by erythrocyte hemolysis test and skin corrosion resistance test in rats.Based on the linear regression analysis of the experimental data of erythrocyte hemolysis,the linear regression equation of the concentration of Buddleja refining solution X and the hemolysis rate Y is Y=1.0945X+0.0347,R2=0.9958.When the concentration was 1.0 g/mL,the hemolysis rate was1.104%<5%.After the resistance experiment,the value is about 12 between the values that meet the specified requirements.After the SRB staining experiment,through absorption comparison,it is preliminarily concluded that the refined liquid does not have skin irritation and corrosion in the result of this experiment.Through the tyrosinase activity test in vitro,from the perspective of monophenolase and diphenolase,the effects of the tested solution on the enzymatic mechanism were discussed.It was found that the biochemical enzymatic activity of Buddleja refining liquid to inhibit monophenolase activity was low,and the activity of p-diphenolase was high,IC50=305.007 mg/mL;the concentration of Buddleja refining liquid was positive to the activity of inhibiting tyrosinase in biochemical enzymes.Related.That is to say,the inhibition of the diphenolase activity of Buddleja refining liquid and L-dopa in enzymatic research is reversible competitive inhibition.According to the enzymatic formula,the inhibition constants of[EI]and[ESI]are KI=0.0048 mg/mL,KIS=0.0099 mg/mL,and the KIS value is greater than KI,that is,the refined liquid can have better enzyme inhibition effect.Based on the laboratory experience formula,the stability,sensory properties and skin irritation were taken as indicators to determine the best formula of Buddleja.Determine the best emulsifier is sodium lauryl alcohol phosphate(HR-S2),the best thickener is magnesium lithium silicate,the best addition amount of refined liquid is 2.5%,which has the best comprehensive effect;continue to conduct response surface experiments optimized,it is concluded that the addition amount of thickener,emulsifier and refined liquid are 1.6%,2.5%and 2.6%respectively;and the stability test of the dosage form has been carried out,and the inspection of centrifugation,alternating cold and heat,and heat and cold resistance has been completed.With reference to national standards,human patch experiments and irritation experiments were carried out,and it was preliminarily verified that Buddleja cream applied to the skin would not cause adverse reactions.Taking the reference substance of buddleoside and luteolin as control,the contents of buddleoside and luteolin in buddleoside cream were determined by HPLC,and the method was investigated.The average content of buddleoside and luteolin was 0.052 mg/g and 0.105 mg/g,respectively,So as to study the quality standard of Buddleja inebriate cream. |
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