High Enrichment and Purification Process of Active Components from the Aerial Parts of Gnaphalium Affine
An HPLC method was established for the determination and quantification of 5 representative compounds,namely chlorogenic acid,caffeic acid,4,5-dicaffeoylquinic acid,1,3,4-tricaffeoylquinic acid and luteolin,in the resin separation processes.The optimal resin was selected by the static and dynamic adsorption/desorption experiments.Process optimization of packed column separation was conducted.A UPLC method was also established for the determination and quantification of 10 representative compounds,e.g.,chlorogenic acid,caffeic acid,caffeoylquinic acids and flavonoids,in alkali precipitation processes.The purities and recovery ratios of active compounds were determined to optimize the alkali precipitation process parameters,including solvent type,solvent addition sequence,material-liquid ratio and amount of alkali.The results showed that LS305 resin had better adsorption and desorption capacities for the five components.The majority of impurities could be washed by 0.5％formic acid,which could remove about 82％of impurities and solids.The efficient separation of active compounds could be achieved by 60％aqueous ethanol.The alkali precipitation was carried out with ethyl acetate-ethanol(4∶1)as the precipitant in a material-liquid ratio of 100 mg∶1 mL.Then,0.15％ammonia was added.After filtration and evaporation,the novel extract of Gnaphalium affine was obtained with the total contents(mass fraction)of 10 compounds more than 50％.
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