Study on Ultrasonic Assisted Enzyme Extraction,Aqueous Two-Phase Extraction and Macroporous Resin Purification of Gastrodin
Objective:To optimize the ultrasonic-assisted enzyme extraction of gastrodin from Tianma(Gastrodia elata)and further purify it according to aqueous two-phase extraction system and macroporous resin,so as to provide process reference for refining high-purity gastrodin.Methods:The optimum conditions of ultrasonic-assisted enzyme extraction were obtained by single factor and orthogonal optimization.The optimum conditions of aqueous two-phase extraction were obtained by drawing the two-phase diagram and process investigation.The optimum conditions of macroporous resin were obtained by process investigation,with optimization of loading and elution conditions.Results:The appropriate conditions were α-amylase enzymes,1:30 of feed liquid,0.8%of enzymatic quantity,80 min of enzymolysis time,60℃of enzymolysis temperature,and pH 5.5 for enzyme solution.The optimal conditions for aqueous two-phase extraction were ethanol volume fraction of 47.5%,ammonium sulfate mass fraction of 15.0%,extraction liquid concentration of 0.2 g/mL,and suitable for macroporous resin screening as HPD100.The optimal sampling conditions were sampling concentration of 0.2 g/mL,with diameter to height ratio of 1:5.The loading flow rate was 3 BV/h,and the loading volume was 100 mL.The optimal elution condition was the integral number of elution liquid 30.0%,with the elution flow rate of 4 BV/h,and the elution volume was 230 mL.Conclusion:In this study,the optimization results of the ultrasonic-assisted enzyme extraction process were good.The purity of gastrodin was greatly improved by aqueous two-phase system and macroporous resin purification.And this series of processes is stable and reliable,laying the foundation for subsequent preparations and new drug research and development.
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