乙醇-硫酸铵双水相体系萃取苹果多酚及与血清胆红素相关性研究

Ethanol-ammonium Sulfate Aqueous Two Phase System Extraction of Apple Polyphenol and Association with Serum Bilirubin

【目的】建立乙醇-硫酸铵的双水相体系,萃取苹果中的多酚;探讨苹果多酚与血清胆红素的相关性.【方法】以萃取率和分配系数为指标,通过单因素实验,考察硫酸铵用量,乙醇浓度,温度对乙醇硫酸铵双水相体系分配行为的影响.采用响应曲面法进一步优化获得最佳萃取条件.选取15例高胆红素患者的血清,将在最佳萃取条件下萃取的苹果多酚以1:1比例与血清混合,体外37℃孵化,在不同时间测定血清中总胆红素、结合胆红素的浓度,分析苹果多酚与血清胆红素的相关性.【结果】乙醇硫酸铵的双水相体系为:硫酸铵3.1g,70%乙醇,温度为40℃,萃取率达可到98.89%,苹果多酚对未结合胆红素、总胆红素浓度呈正相关(r分别为0.994~0.996,0.610~0.993,P均小于<0.05),结合胆红素浓度无明显的相关性(r为0.335~0.662,P>0.05)随着苹果多酚浓度的增加,降低趋势变化较快.【结论】构建的乙醇硫酸铵双水相体系可以较好的萃取苹果多酚,方法绿色,易操作;苹果多酚可降低血清中总胆红素、未结合胆红素的浓度.

Objective:Establish ethanol-ammonium sulphate aqueous two-phase system to extract apple polyphenols and to explore the correlation between apple polyphenols and the serum bilirubin.Methods:The extraction rate and distribution coefficient were taken as the index.Through the single factor experiment,the dosage of ammonium sulfate,ethanol concentration and the temperature were investigated to research distribution behavior of double water phase system.Response surface method is used to further optimize the related parameters to obtain the optimum extraction conditions.The serum which was selected 15 hyperbilirubinemia patients was mixed with the extracted apple.The apple concentration of total bilirubin and direct bilirubin in serum was determined at different time to analyze the correlation of apple polyphenol with serum bilirubin.Results:Aqueous two-phase extraction of apple polyphenol the best conditions for:ammonium sulfate dosage is 3.1g,ethanol concentration is 70%,temperature 40℃,extraction rate of98.89%.There is significant positive association between the apple polyphenol and the indirect bilirubin and the total bilirubin(r is respective 0.994~0.996 and 0.610~0.993,P is less than 0.05),which were a decreasing trend.The direct bilirubin shows no significant association(r is 0.335~0.662,P is more than 0.05).Conclusion:Ethanol ammonium sulphate aqueous two-phase system can better extract the apple polyphenol.The determined method was green and easy to operate;Apple polyphenols could reduce concentration of total bilirubin and indirect bilirubin in the serum.