Preparation And Preliminary Evaluation Of Plural Emulsion Gel For Anti-vitiligo

The Bai Fu Ning is a drug that has effect on anti-vitiligo in this study.It is a plural emulsion gel and its functional component is the effective part of the wild Yang Jiaowan from Weibei for anti-vitiligo.Yang Jiaowan is a mutant of Metaplexis japonica(Thunb.)Mak.of Metaplexis Asclepiadaceae.In Weibei folk,the white juice of Yang Jiaowan could be used to treat vitiligo,and the same effect of the same species plant was recorded in “Guang Jifang”.Yang Jiaowan,the grass plant resources are very rich,but it has been run its course for many years.In order to develop and utilize this natural resource to explore new drug for the treatment of stubborn vitiligo,Yang Jiaowan was researched in this paper.The research contents included four aspects: selection of the effective part for anti-vitiligo of Yangjiaowan,component test,preparation process,preparation and preliminary evaluation of dosage forms.Firstly,the chemical constituents of Yang Jiaowan were systematically predicted by chemical methods.According to the result,the monomer compounds were obtained by phytochemical research method,and the structures were identified.Secondly,integrating the study results,the constituents were divided into four categories according to the polarity,and different extracts were obtained with four different polar solvents.The effective part of Yang Jiaowan was screened by Bioactivity Guided Isolation method(BGI).Thirdly,the optimization of the extraction process of the effective part was carried out by response surface experiment(4 factors and 3 levels).Finally,the preparation process was studied by means of experiment comparison and comprehensive evaluation,and the pharmacodynamics of the Bai Fu Ning was preliminarly evaluated by measuring the biological activity.The main resluts were as follows:(1)Selection of the effective part of Yang Jiaowan for anti-vitiligoYang Jiaowan contained 14 kinds of chemical constituents by systematic prediction and qualitative identification,such as saponins,steroids,phytosterols,flavonoids,alkaloids,organic acids,etc.Eight different polarity extractsincluding the petroleum ether extract,ethyl acetate extract,n-butanol extract,water extraction and alcohol precipitation extract(20%,40%,60%,80%,95%)were determined by the tyrosinase activity measurement method which was established and validated.The result showed that n-butanol extract had the most significant effect on the activation of tyrosinase among them,and the activation rate was 30.71%.At the same time,psoralen as the positive control,and its activation rate was 39.34% in the experimental range.(2)Phytochemistry research of the effective part of Yang JiaowanSaponins were detected in n-butanol extract by physical and chemical properties test,and two saponins spots were observed by thin layer chromatography.Four monomer compounds were obtained from n-butanol extract by the column chromatography separation,crystallization purification and other processing.Their structures were identified as weigoside(compound1),?-sitosterol(compound 2),robinin(Kaempferol-3-O-glucosyl-rhamnosyl-7-O-rhamnoside,compound 3)and inositol(compound 4)in terms of physical and chemical properties,chromatography,spectroscopy and other methods.(3)Extraction process of the effective part of Yang JiaowanThe stem structure of Yang Jiaowan was composed of epidermal parenchyma cells,cortical parenchyma cells,secondary phloem parenchyma cells,secondary xylem parenchyma cells and myeloid cells by microscopic observation.The leaf structure of Yang Jiaowan consist of epidermal parenchyma cells and mesophyll cells by the above method.Histochemical localization results showed that saponins were mainly distributed in parenchyma cells,secondary phloem parenchyma cells and myeloid cells of the stem,and saponins in mesophyll cells of the leaf.Extraction process: Based on the results of single factor experiment,a response surface methodology was designed to optimize the extraction process with the total saponin labeled content as the indicator.The results of the optimum extraction conditions were as follows: ethanol concentration was 76%,liquid-solid ratio was 10.32:1(mL/g),reflow temperature was 70.2 ?,and reflow time was 2.52 h in the experimental range.According to the optimum result,the experiment was carried out 5 times repeatly.The yield of solid content was 4.32%,and the labeled content of the total saponin was 60.39%.It wasverified that the relative standard deviation(RSD)of the process was 1.5%.(4)Preparation and preliminary evaluation of the pharmacodynamics of the Bai Fu NingThe physical and chemical properties of n-butanol extract regarded as gist,the clinical applicability as criterion,w/o/w plural emulsion gel was more suitable for the product by literature investigation and contrastive analysis for six external formulations.The formulation and molding process parameters were optimized by L9(34)orthogonal experiment,and the result was that prescription compositions were1.0 g drug,1.2 g glyceryl monostearate,0.5 g white petrolatum,2.7 g liquid paraffin,0.25 g stearic acid,0.2 g plate-80,0.1 gTween-80,0.1 g CMC-Na,0.1 g PVA124,a small amount of additives and appropriate amount of distilled water.Then the formulation compositions were emulsified at 80? to prepare product.The result showed that the product colour was beige,and with good stability.The labeled amount of functional component(robinin as indicator)was no less than8.0% in the product.The drug release rate was 75.6% in 0~8h by drug-releasing experiment in vitro,and the correlation coefficient of the first-order model was higher.The experimental results of preliminary evaluation of the pharmacodynamics of the Bai Fu Ning showed that the product had no stimulation to skin,and it had an effect on the activation of tyrosinase,the activation rate was 30.62%.At the same time,psoralen tincture as the positive control,the activation rate was 57.34%.Therefore,the results could be concluded as follows: the effective ingredient in effective part of the wild Yang Jiaowan from Weibei for anti-vitiligo was glycosides.The labeled amount of the total saponins was no less than 60% in the effective part.Weigoside,?-sitosterol,robinin,inositol and two kinds of saponins were purified from the effective part of Yang Jiaowan,and inositol was first found in the same species plant.The process of using ethanol and n-butanol extraction was stable,and with strong operation.The activated effect that plural emulsion gel acted on tyrosinase was reliable.The results of this study could provide a reliable experimental basis for the development of new drug for the treatment of vitiligo.