葡聚糖凝胶柱层析富集黑果枸杞总花色苷工艺及其抗氧化性研究

以黑果枸杞花色苷提取物为原料,研究Sephadex LH-20葡聚糖凝胶富集纯化花色苷工艺条件,分别考察不同上样液浓度、p H值和上样流速对花色苷的吸附率影响,同时考察不同洗脱液浓度、p H值和洗脱流速对花色苷的解吸效果,并比较不同产物的总花色苷含量和抗氧化性。通过单因素与正交试验确定最佳富集纯化工艺条件为:质量浓度25 mg/L、p H值为3.5的样品溶液,以1.0 mL/min流速上样至Sephadex LH-20葡聚糖凝胶柱后,采用85%、p H值为3.5的乙醇溶液,以1.0 mL/min流速洗脱,对总花色苷的吸附率和解吸率分别为90.9%、85.3%。通过定量分析可知,凝胶柱层析富集后产物的总花色苷含量高于树脂纯化物,同时抗氧化性试验结果表明,对两种自由基的清除能力顺序依次为凝胶纯化>树脂纯化>提取物。

Study of Enrichment Process of Total Anthocyanins from Lycium ruthenicum Murr.by Sephadex LH-20 and Its Antioxidant Activity

The enrichment process of total anthocyanins crude extract from Lycium ruthenicum Murr.with sephadex LH-20 was investigated.The influence of target object adsorption of different sample concentration,adsorption flow rate,and sample p H value were reseached.At the same time,the influence of anthocyanins recovery of different ethyl alcohol concentration,elution flow rate,and eluent p H value were investigated.The best purification conditions by single factor and orthogonal test showed that pH 3.5 sample solution with anthocyanins concentration of 25 mg/L,adsorption velocity 1.0 m L/min,eluent ethanol concentration of 85%,eluent pH of 3.5,elution velocity of 1.0 mL/min,the adsorption rate and desorption rate of anthocyanins was 90.9%,85.3%respectively.The total anthocyanins content of sample after purification was obviously higher than the resin purification product,Moreover,the results from two antioxidant assays consistently indicated that the antioxidant capacity of product by gel purification,resin purification and microwave-assisted revealed a decreasing trend in turn.