Aqueous Two-Phase Extraction And Purification Of Phloridzin From Malus Micromalus

Malus micromalus is a local characteristics fruit,which enriched nutrition and functional material,but at present the study of Malus spectabilis was more tend to products processing,the study of functional components in the fruit is still inadequate.In this paper,ultrasonic-assisted aqueous two-phase extraction technology was used to extract phloridzin from Malus micromalus fruit,and optimized the technological parameters of extraction as well as compared with traditional method;Then,the crude extracts of phloridzin was purifed using macroporous adsorption resin,the response surface method was used to determine the optimum conditions of adsorption and desorption according to the results of single factor experiment.Meanwhile,the antioxidant activity?scavenging ability of DPPH· and ABTS·+,the total reducing power?of phloridzin refined material eluted by different concentrations of ethanol was investigated.The main results were followed:?1?The recovery?R₁?and partition coefficient?K?of phloridzin as the response value,the optimum technological conditions of ultrasonic-assisted aqueous two-phase extraction was determined by the single factor experiments and response surface method: 35% mass fraction of ethanol,16% mass fraction of ammonium sulfate,57 of liquid-solid ratio,extraction temperature was 36 ?.According to this condition,the recovery?R₁?of and partition coefficient?K?of phloridzin were 92.86 ± 0.078% and 6.55 ± 0.08,which is no obvious difference compared with theoretical value?92.94%? 6.59,P>0.05?,indicated that aqueous two-phase extraction was a effective method for phloridzin extracted from Malus micromalus fruit.?2?Aqueous two-phase technique compared with traditional organic solvent extraction method?35%?w/w?ethanol and 80%?v/v?etanol extranction method?,not only have higher extraction quantity of phloridzin,but also can remove much more sugar?84.32% and 65.27%?and protein]?91.63% and 34.62%?,and keep its antioxidant activity.?3?The adsorption rate of ADS-7,AB-8,DM-2 and D101 resin for phloridzin were all over 63%,of which the adsorption rate of ADS-7 resin was highest?79%?.That means these four resins has good adsorption properties for phloridzin in 7 kinds of macroporous resin?NKA-9,AB-8,D101,ADS-17,the ADS-F8,ADS-7,DM-2?.?4?AB-8,DM-2,D101 and NKA-9 resin has better desorption properties for phloridzin extracted from Malus micromalus fruit,the desorption rate of AB-8 resin was highest,at 93.28%.And the ADS-7 resin has the highest adsorption rate,but the desorption rate was low,only 31.25%.AB-8,DM-2 and D101 resin can fast adsorb the phloridzin,and the adsorption and desorption rates of AB-8 resin for phloridzin were all the largest in.these three resins Therefore,the AB-8 resin was suitable for purification of phloridzin.?5?The optimum parameters of purification of phloridzin by AB-B macroporous resin was determined by response surface method.The adsorption conditions: adsorption rate of 1 mL/min,the sample solution pH value of 1.5,the sample liquid concentration of 0.12 mg/mL;The desorption conditions: ethanol concentration was 70%,the eluent pH value of 6.7,the elution rate of 2 ml/min.According to the experiment of these conditions,the adsorption and desorption rates of phloridzin were 90.89±0.02% and 90.89±0.37%,which was no obvious difference compared with theoretical value?90.17%? 95.69%?.?6?The study of in vitro antioxidant activity of phloridzin refined material eluted by different concentrations of ethanol and crude extracts was found that the phloridzin refined material eluted by 70% ethanol has the highest scavenging capacity of DPPH and ABTS free radical and total reducing power,which was followed by 90% ethanol eluate,50% ethanol eluate and 30% ethanol eluate,and were higher than that of phloridzin crude extracts by aqueous two-phase extraction from Malus micromalus fruit.Meanwhile,the content of phloridzin in 70% ethanol eluate reached 37.89?g/mL,higher than the content of crude phloridzin extracts 44 times,combined the antioxidant activity results show that the method of macroporous resin for preliminary refined of phloridzin is meaningful and effective.