目的:建立AB-8大孔树脂纯化稠李花色苷的方法,并检测稠李花色苷体外抗氧化活性。方法:通过AB-8大孔树脂对稠李花色苷的动态吸附与解吸条件优化,确定最佳纯化条件;采用四唑氮蓝(nitroblue tetrazolium,NBT)光化还原法测定花色苷体外抗氧化活性,2’,7’-二氢二氯荧光素二乙酸酯(2’,7’-dichlorodihydrofluoresce in diacetate,DCFH-DA)法观察花色苷对H2O2诱导的N2A细胞氧化损伤的保护作用。结果:稠李花色苷的最佳动态纯化条件是:吸附流速0.01 m L/s,粗提液质量浓度8 mg/m L,过柱次数2次;解吸剂乙醇体积分数70%,流速0.01 m L/s,pH 3,纯化之后稠李花色苷的色价为57.1,纯化了16.31倍;NBT实验结果表明,稠李花色苷具有清除超氧阴离子自由基的能力,且具有较好的剂量-效应关系;0.025 mg/m L的稠李花色苷能够保护H2O2损伤的N2A细胞,0.05 mg/m L的稠李花色苷能够降低细胞内活性氧水平。结论:AB-8大孔树脂是纯化稠李花色苷的一种有效方法,稠李花色苷在一定质量浓度范围内具有较好的体外抗氧化活性。
Purpose: To establish the best purification conditions for anthocyanins from the fruit pulp of Padus racemosa with AB-8 macropomus resin and detect the in vitro antioxidant activity of the extracted anthocyanins. Methods: The adsorption and desorption properties of macroporous resin AB-8 for anthocyanins from Padus racemosa were investigated to optimize the purification procedure, and the antioxidant activity in vitro was detected by nitroblue tetrazolium (NBT) photochemical reduction method. The protective effect of the extracted anthocyanins against oxidative stress induced by H202 in N2A cells was analyzed by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe assay. Results: The best purification conditions were achieved by adsorption performed twice with at a flow rate of 0.01 mL/s after adjustment of the crude extract concentration to 8 mg/mL and subsequent desorption with 70% ethanol acidified to pH 3 at a flow rate of 0.01 mL/s. The color value after purification was 57.1, achieving a 16.31-fold purification factor. The anthocyanins had a strong superoxide anion radical scavenging capacity in a dose-dependent manner. N2A cells could be protected by the anthocyanins at a concentration of 0.025 mg/mL, and the level of reactive oxide species in N2A cells could be reduced by the anthocyanins at 0.05 mg/mL. Conclusion: Macroporous resin AB-8 is an effective adsorbent for the purification of Padus racemosa anthoeyanins, and Padus racemosa anthocyanins have strong antioxidant activity in v/tro in a certain concentration range.