星点设计-效应面法优化大孔树脂纯化还原型萝卜硫苷工艺及体外抗氧化活性研究

Process study on macroporous resin for purification of glucoraphasatin by central composite design-response surface methodology and antioxidant activity of glucoraphasatin in vitro

为研究大孔树脂纯化还原型萝卜硫苷的最佳工艺及其抗氧化能力。实验以8种不同型号的大孔树脂对还原型萝卜硫苷的比吸附量、吸附率和洗脱率为指标筛选出最佳型号的大孔树脂,采用单因素考察和星点设计-效应面法优选出大孔树脂纯化还原型萝卜硫苷的工艺参数,通过测定纯化前后还原型萝卜硫苷提取物对DPPH自由基及ABTS+·自由基的清除能力来表征其抗氧化活性。结果表明,HPD-722型大孔树脂纯化还原型萝卜硫苷效果最好,最佳纯化工艺为:上样液pH 5.3,上样流速2.5 BV/h,上样液浓度0.53 mg/mL;洗脱液为70%乙醇溶液,洗脱液体积为2.5 BV,洗脱液流速为1.5 BV/h,还原型萝卜硫苷纯度由0.404%提高到17.903%,纯度提高了44.35倍,纯化后的还原型萝卜硫苷GRH提取物与萝卜提取液相比,清除DPPH自由基和ABTS+·自由基的能力分别提高了67.31和45.27倍。

This study was to investigate the optimal purification process for macroporous resins of glucoraphasatin and the antioxidation.The best type of macroporous resin was selected by 8 different types of macroporous resins for the specific adsorption amounts,adsorption and elution rates of reduced glucoraphasatin,the purification technology for glucoraphasatin was optimized by single factor investigation and central composite design-response surface methodology.And the antioxidant activity of glucoraphasatin extractive purification before and after was characterized by the scavenging capacity of DPPH and ABTS+·free radicals.The results show that HPD-722 macroporous resin has the best effect on purifying glucoraphasatin,the optimal condition was as follows as the sample solution pH is 5.3,flow rate is 2.5 BV/h,the concentration is 0.53 mg/mL,the eluent was 70%ethanol solution,the eluent volume was 2.5 BV and the flow rate was 1.5 BV/h.The purity increased by 44.35 times from 0.404%to 17.903%.Compared with the radish extract,the ability of the purified glucoraphasatin extractive to scavenge DPPH free radicals and ABTS+ free radicals increased by 67.31 and 45.27 times,respectively.