大孔树脂分离纯化毛酸浆叶黄素及其抗氧化活性评价

Separation and Purification of Lutein from Physalis pubescens by Macroporous Resin and Evaluation of Its Antioxidant Activity

目的:研究大孔树脂分离纯化毛酸浆叶黄素的工艺条件,提高毛酸浆叶黄素的抗氧化活性。方法:在单因素实验的基础上,采用正交实验优化大孔树脂HP-20静态吸附毛酸浆叶黄素的工艺条件;以解析率为参考指标,探究毛酸浆叶黄素动态解析的工艺条件;通过测定毛酸浆叶黄素对OH自由基和DPPH自由基的清除率,评价其纯化前后的抗氧化活性。结果:静态吸附叶黄素最佳条件为:HP-20型树脂与叶黄素粗提液比例3∶1(g/mL)、叶黄素浓度400 mg/mL、吸附温度20℃、吸附时间4 h,此条件下吸附率达到63.44%。动态解析叶黄素最佳条件为:洗脱流速0.42 mL/min、洗脱剂60%乙醇溶液、上样液质量浓度800 mg/mL,此时解析率为56.16%。体外抗氧化研究结果显示,纯化前后叶黄素对OH自由基的最高清除率分别是97.66%和98.33%,对DPPH自由基的最高清除率分别是88.21%和90.47%。可见,纯化后的叶黄素能更好地清除OH自由基和DPPH自由基,且对OH自由基的清除效果更强。结论:优选大孔树脂分离纯化毛酸浆叶黄素工艺条件合理可行,且纯化后的叶黄素具有更强的抗氧化活性。

Objective:Purification process conditions of lutein in Physalis pubescens by macroporous resin were studied to improve the antioxidant activity of lutein in P.pubescens.Methods:On the basis of single factor experiments,orthogonal experiments were used to optimize the process conditions for static adsorption of lutein in P.pubescens by macroporous resin HP-20.At the same time,with desorption rate as the evaluation index,the dynamic analysis process conditions of lutein in P.pubescens was explored,the antioxidant activity of lutein in P.pubescens before and after purification was evaluated through measuring its scavenging rates against OH and DPPH radicals.Results:The optimal static adsorption conditions were as follows:a ratio of HP-20 resin to crude lutein extract of 3∶1,lutein concentration of 400 mg/mL,adsorption temperature of 20℃,adsorption time of 4 h,resulting in an adsorption rate of 63.44%.The optimal dynamic analysis conditions were as follows:elution flow rate of 0.42 mL/min,eluent ethanol concentration of 60%,sample concentration of 800 mg/mL,achieving an desorption rate of 56.16%.The results of in vitro antioxidant studies showed that the highest scavenging rates of lutein before and after purification against OH radicals were 97.66%and 98.33%,respectively,while those against DPPH radicals were 88.21%and 90.47%,respectively.It could be seen that purified lutein could better scavenge OH and DPPH radicals,and had a stronger scavenging effect on OH radicals.Conclusion:The optimized process conditions for separating and purifying lutein in P.pubescens by macroporous resin was reasonable and feasible,and the purified lutein has strong antioxidant activity.