Study On Separation And Purification Of Madecassoside From Centella Asiatica And Preparation Of Scald Spray

Centella asiatica is rich in triterpenoids,most of which are madecassoside.Guizhou is rich in Centella asiatica.In order to make full use of Centella asiatica,this article used Centella asiatica as raw material.First,the determination method of madecassoside was determined;Secondly,the best extraction process was determined by the L₉(3⁴)orthogonal test;Then,the madecassoside that the content of more than 95%was prepared by macroporous adsorption resin,alumina,crystallization and preparative HPLC methods;Finally,the scald spray with Centella asiatica extract were prepared,conducted stability studies and drafted quality standards.The main research contents were as follows:1.Determination method of madecassoside in Centella asiatica:By consulting related literature and conducting methodological investigations,the content detection method of madecassoside HPLC was optimized:Agilent 1260,chromatographic column:Neptune C18 analytical column(250 mm×4.6 mm,5?m),mobile phase:Acetonitrile-0.2%phosphoric acid water(0 min 25:75?20 min 25:75?30 min30:70?40 min 40:60?50 min 50:50),detection wavelength:206nm,column temperature:25?,flow rate:1.0 m L/min,Injection volume:10?L.The negative control indicated that the excipients had no interference in the determination of the content of madecassoside in the spray.the determination method of madecassoside content in Centella asiatica was suitable for the determination of the content of scald spray.2.Study on the extraction technology of madecassoside in Centella asiatica:By using L₉(3⁴)orthogonal test to determine the best extraction process.The optimal extraction process parameter was ethanol concentration of 50%,the material-to-liquid ratio was 1:20,the extraction temperature was 30°C,the extraction time was two hours,and the extraction time was one time.Under these conditions,the extraction amount of madecassoside was 5.63 mg/g,and the extraction rate was88.33%.3.Study on the purification process of madecassoside in Centella asiatica:First,the parameters for purification of madecassoside by macroporous adsorption resin were as follows:the loading amount was 19.8 mg/g,the loading flow rate was 1BV/h,the ethanol elution volume fraction was 50%,the ethanol elution volume was4 BV,the resin diameter-to-height ratio was 1:6.Under these conditions,the content of madecassoside was 27.37%.Secondly,it was determined that alumina was used for decolorization and purification,and the decolorization process parameters were as follows:The chromatographic reagent was 100-200 mesh acidic alumina,the elution solvent was 10%ethanol,the elution volume was 3 BV,and the ratio of the weight of Centella asiatica extract to the amount of acidic alumina was 1:10.Then,a pilot study on the extraction and purification of madecassoside was carried out.The results showed that the content of madecassoside in the obtained Centella asiatica extract was more than 54%,and the yield was more than 2.24%.The recovery rate was more than 63.57%.The results of three batches proved that the process was repeatable,and the product quality was relatively stable.Finally,the crystallization method was used for purification,the crystallization solvent was 50%methanol,and the crystallization-to-liquid ratio was 1:5.The purity of madecassoside by crystallizating was more than 85%.The purity of madecassoside which were separated by preparative high performance liquid chromatography was more than98%.4.Research on the preparation process of scald spray:Through researching and selecting the auxiliary materials,the final formulation and preparation process of the scald spray were determined as follows:1.0 g Centella asiatica extract added to 10 g60%ethanol and shook to dissolve,added to 0.5 g Tween-80,6 g 1,2-propanediol and 0.2 g benzoic acid,stiring,added sandalwood essence,added distilled water to100 m L,filtering,added sodium hydroxide to adjust the p H to 4.5?6.5,shaking,sterilizing,packing.Researching the quality standards for sprays.Three batches of scald spray were subjected to high temperature,high humidity and light tests.The data showed that high temperature and high humidity had no effect on scald spray,but light would reduce the content of madecassoside and increase the content of asiaticoside.Therefore,the scald spray were packaged in light-proof.The accelerated stability test results showed that these inspection items had no obvious changes,and the stability was good.Therefore,it was recommended that the scald spray should be stored at protected from light,and sealed.The storage time was one year.This experiment not only studied a continuous extraction and purification route of madecassoside,but also prepared scald spray which were stable and feasible.The optimized HPLC detection method of madecassoside was better separated from asiaticoside-B.The extraction,separation and purification process was continuous and simple to operation,and the scald spray had good stability.