The Production Of Total Sapindus-saponins From Sapindus Mukurossi Gaertn

The pericarp of sapindus has a large number of saponins which are mainly Oleanolic acidsaponins and are called the total sapindus-saponins(TSS).TSS is a natural surfactant and hasbiological activities such as antipruritic and sterilization.TSS of high purity was made as qualitycontrol with the column seperation.The process of water-extraction and microbial fermentation wasadopted to separate and purify TSS.The results were shown as follows.1With the method of ethanol extraction,butanol extraction,AB-8macroporous resin and MCIGel column separation,a high purity of more than95%of TSS was made as quality control.Bubbleexpriments proved that TSS produced honeycomb foam,lasting15min;Libermann-Buehard reactionshowed the color changes:yellow-red-purple-blue;Hemolysis test proved hemolytic.Thecomponents named SapindosideB(882),SapindosideA(750),Mukurozi-saponinE1(924),SapindosideL(924),Mukurozi-saponin G(966),Sapindoside M(966) were analysed by HPLC/MS,which wereidentified as Oleanolic acid saponins.Oleanolic acid was chose as standard.Sapogenin of TSS afterhydrolysis has a maximum absorption at550nm with the method of vanillin–sulfuric acid,whichwas the same as Oleanolic acid in the condition of35%hydrochloric acid,high temperature andpressure and time of2h,the yield of sapogenin were the highest.The purity of TSS were calculatedto more than95.2%.It can be further conclued that TSS were mainly oleanolic acid saponins andcan be used for quality control as standard.2The standard of TSS had a greatest absorption at478nm in the method of vanillin–sulfuricacid.The liner regression equation at a range of100μg~1000μg was y=0.0006x+0.0838.r=0.9996,which showed a good linear relationship and could be used as standard curve of TSS.Thismethod had a high precision and repeatance with the average recovery of99.83%,which provided abasis method for quality control of TSS.3Aspergillus oryzae fementation was the most effective to purify TSS than natural fementationand yeast fementation.Water extraction of sterilization by Aspergillus oryzae fementation wasalmost equal to that of no sterilization.Aspergillus oryzae fementation was selected to purify TSS ofwater extraction of no sterilization.4The optimum water extraction conditions were as follows:three times extraction with waterunder40℃,sample/liquid ratio1:7,1:3,1:2, and5h,2h,1h spent.The yield of TSS was33.34%and the purity48.71%.The best Aspergillus oryzae fermentation conditions were as follows:waterextraction of Sapindus mukorossi Gaertn not sterile,temperature30℃, vaccination age12h,inoculation3%, shaking speed150r/min,fermentation time7d.In these conditions,the yield of TSSdecreased slightly（33.34%→29.89%）,and the purity increased from48.31%to82.47%.When pHvalue reached about7.6,the purity of TSS by Aspergillus oryzae fermentation reached the largest.So, the pH value was used as the index of the end point of fermentation for purification.