Optimization Of The Extraction Process Route For The Glycosides In Albizia Julibrissin And Establishment Of Its Quality Standard

The color property of UV spectrophotometry was poor and it was susceptible toenvironment, equipment and other external factors. Therefore, the establishment of aspecific high performance liquid chromatography to determine the total saponinscontent of Albizia julibrissin was of great significance. With the HPLC method, aprocess route to get the highest total saponins was accessed by orthogonal test of fourfactors and three levels. With Albizia julibrissin containing complex compositions,this paper aimed to get several components of high activity and relatively simplecomposition, and calibrated a few peaks as fingerprint peaks, thus an effective qualitystandards could be services to the determination of the total saponins content.The establishment of the HPLC quantitative analysis method to determinate thecontent of total saponins of effective anti-neovascularization from Albizia julibrissinprovided a basis for controlling the quality in Albizia julibrissin. As the mothernucleus of most Albizia saponins came out to be oleanolic acid, which was thesapogenin attained by acid hydrolysis of saponins. On this base, the HPLC methodwas established. In order to find the best hydrolysis of saponins in Albizia julibrissin,each factor’s affection on the degree of hydroysis respectively was studied, includingthe kind and amout of acid, reaction time, acid concentration and reaction temperature,by doing single fator(factor) and orthogonal experiments. The HPLC method wascompared with UV spectrophotometry in their specificity. The best kind of acid forhydroplysis of saponins was H2SO4; and the maximum influence factor was reactiontime, then was the amount of acid, acid concentration and reaction temperature;3mol/L H2SO4, liquid to material ratio1:150,85℃and4h reaction time came out tobe the best condition of hydrolysis. Compared with UV spectrophotometry, theoleanolic acid obtained from Albizia julibrissin was determined by this method withgood precision, sensitivity and specificity.According to the established method of detecting total saponins content, thisexperiment armed to find a process route of highest extraction efficiency throughorthogonal test of four factors and three levels,and got component1to9. each factor’saffection on the degree of the route respectively was studied, including the ethanolconcentration, reflux time, extraction times, extraction. the maximum influence factorwas extract times, then was the the number of extraction,reflux time and ethanolconcentration.70%ethanol for2times, reflux3h, extracting4times with water-saturated n-butanol came out to be the best process route of extraction.In this paper, the component A and B were obtained from the water-saturatedn-butanol extraction phase of Albizia julibrissin by SPE technology.With HMEC-1evaluating the activity of the component A and B, the component A was hardly active,while the IC50of component B was corresponding to1.93±0.07μg/mL. The averageextraction rate of component B was52.6%with three times repeatedly. Then thecomponent B-1,B-2,B-3,B-4and B-5were obtained from the component B bypreparative chromatography technology. With HMEC-1evaluating the activity of thecomponents, the IC50of component B-4was corresponding to1.449±0.11μg/mL.Finally, component B-4was analyzed by HPLC, and the results showed that therewere three peaks of best separation and biggest peak area. The three peaks werecalibrated as the ingerprint peaks in the chromatogram in the fingerprint study ofAlbizia. These three peaks correspond to the14,21and22peak of the entirechromatogram,so calibrated the14,21and22peak for the characteristic peaks anddetected the peak area ratio of three characteristic peaks respectively,The resultsshow:the area of14peak less than25.7mAU*min;the area of21peak less than55.89mAU*min;the area of22peak less than23.79mAU*min.