The Integration Technology Of Extraction And Separation For Proanthocyanidins From Black Wattle Bark

In this subject ultrasonic-assisted extraction, macroporous resin adsorption andhigh performace liquid preparative chromatography were coupled to extract andseparate proanthocyanidins from Black Wattle Bark. the results are summarized asfollows:Response surface method was used to optimize the process of ultrasonicassisted extraction of proanthocyanidins from Black Wattle Bark in this experiment,optimization of process parameters based on the extraction solvent50%ethanol areas follows: extract temperature50℃, extraction time33.45min, ultrasonicfrequency67kHz, solid-liquid ratio1:11.0W:V. The average yield ofproanthocyanidins was21.58%under this condition. Under the optimized conditions,the impact of the extraction times was discussed, and the results show that fourextraction times is the best. The total yield of proanthocyanidins can reach to24.26%.The fractionation and purification effects of4kind macroporous adsorbentresins (AB-8, D-101, BS-5, BS-30) on proanthocyanidins from Black Wattle Barkwere investigated,Among the4kinds of macroporous adsorbent resins, AB-8resinhad the most efficient adsorption and desorption properties.by elution withincreasing ethanol concentration and isolation with solvents deposition,thefractionation of proanthocyanidins from Black Wattle Bark were recovered. The totalrecovery rate of proanthocyanidins reach to92.26%.The process conditions of AB-8macroporous adsorption resin to separate andpurify proanthocyanidins from Black Wattle Bark were discussed to determine theoptimum conditions. concentration of elution ethanol solution40%,theconcentration of the sample4mg/ml, the flow rate of sample3ml/min, after theelution of the macroporous resin adsorption,the purity of Proanthocyanidins can beimproved to61.2%.The components and monomer structure of proanthocyanidins from BlackWattle Bark were identified with RP-HPLC/ESI-MS. The results indicate that themain monomers of proanthocyanidins from Black Wattle bark are dimmers andtrimmers. Molecular weight of them are562,578,594and850,866, respectively.Proanthocyanidins were separated by high performance liquid preparativechromatography, the structure of proanthocyanidins were identified with RP-HPLC/ESI-MS. The result indicate that three kinds of dimer wererecovered,Molecular weight and concent of them are562,578,578and9.85%,18.88%,66.84%purity of proanthocyanidins is95.57%.The heavy metals in the crude extract of black wattle bark can be removedeffectively by the method of ethyl acetate extraction, especially lead and copper.Heavy metal test indicates heavy metals contents of the crude extract and the ethylacetate fraction meet the criterion of Green Trade Standards of Importing andExporting Medicinal plants and Preparations.