Study On Extraction Purification And Functional Characteristics Of Anthocyanins From Black Kidney Bean Residue

Black kidney bean（Phaseolus vulgaris Linn.）belongs to the bean family,the staple food is its seeds and pods,the annual herbaceous plant,is one of the beans with a long history of cultivation and edible in China.Through modern analysis technology,black kidney bean contains various functional proteins,various vitamins,a large number of unsaturated fatty acids,flavonoid glycosides,anthocyanins and other functional components.In order to realize the comprehensive utilization of black kidney bean residue,from the black kidney bean processing industry waste.This study selected black kidney bean residue provided by the factory as raw material,through ultrasonic extraction technology,continuous chromatography technology and UPLC-TOF/MS technology,the extraction,separation,purification and composition analysis of black kidney bean anthocyanins from bean residue.Single factor tests were designed to determine the optimum operating parameters in the extraction of black kidney bean anthocyanins.Through the analysis of adsorption capacity and desorption capacity of resin,the most suitable resin was selected.The process parameters of preparation chromatography purification of black kidney bean anthocyanin were optimized.The optimum purification parameters for SMB chromatography were determined.Finally,the stability and antioxidant capacity of the purified anthocyanin solution of black kidney bean were studied,and the anthocyanin powder of black kidney bean was prepared by freeze-drying.The antibacterial activity and cytotoxicity of the anthocyanin powder of black kidney bean were studied.The following conclusions are reached:（1）The optimal extraction parameters of black kidney bean anthocyanins were 70%ethanol concentration,30℃extraction temperature,20 min extraction time and 10.0 m L/g liquid-material ratio.AB-8 resin（adsorption rate 82.58%,desorption rate 78.21%）was the most suitable resin for adsorption and separation of anthocyanins from black soybean residue.In the purification of black kidney bean anthocyanins,the optimal loading solution concentration was200 mg/m L,and the optimal elution flow rate was 2.0 m L/min.By choosing 40%ethanol as the eluent,anthocyanins with higher concentration,higher yield and higher antioxidant activity could be collected(IC₅₀ of DPPH radical scavenging activity was 0.95μg/m L,IC₅₀ of ABTS+·radical scavenging activity was 2.14μg/m L).（2）The optimal operating parameters for the purification and separation of black kidney bean anthocyanins by SMB method are as follows:an adsorption zone flow rate of 12.5 m L/min,refined zone flow rate of 28.5 m L/min,desorbing zone flow rate of 24.5 m L/min,regenerative zone flow rate of 22.5 m L/min,water washing zone flow rate of 38.5 m L/min,and switching time of 1080 s.（3）SMB method is more suitable for industrial production of anthocyanins in black kidney bean than TMB method.The daily sample quantity treated by SMB method（18 L）was significantly higher than that treated by TMB method（3.6 L）（P<0.05）.For unit volume samples,ethanol consumption（3.04 unit volume）and pure water consumption（6.08 unit volume）by SMB method were significantly lower than those by TMB method（6.4 unit volume and 9.6 unit volume）（P<0.05）.There are 3 kinds of anthocyanins in black kidney bean anthocyanins:delphinidin-3-O-glucoside,petunidin-3-O-glucoside and malvidin-3-O-glucoside.（4）In order to improve the preservation rate of black kidney bean anthocyanins,the frequency and degree of heating concentration of anthocyanins should be reduced,and the long-term high-temperature treatment of anthocyanins should be reduced or even avoided,anthocyanins should be stored in a low p H environment.Black kidney bean anthocyanins can inhibit the growth of Staphylococcus aureus and an anti-proliferation effect on B₁₆melanoma cells(IC₅₀=0.846 mg/m L).In conclusion,using simulated moving bed continuous chromatography purification can achieve high efficiency,high material utilization rate,low energy consumption and low-cost separation and purification of black kidney bean anthocyanins from black soybean residue.The anthocyanins obtained from black kidney bean have certain antioxidant,bacteriostatic and cytotoxic properties.This study can provide theoretical reference for the comprehensive utilization of black kidney bean residue,the purification process and industrial utilization of black kidney bean anthocyanins.