曲面响应法优化核桃雄花多酚提取工艺及其组成与抗氧化活性分析

Optimizing polyphenol extraction techniques of walnut male flowers by surface response methodology and its composition and antioxidant activity

  • 摘要: 【目的】采用曲面响应法优化核桃雄花多酚提取工艺,并分析其组成与抗氧化活性,为核桃雄花的综合利用提供技术支撑。【方法】以干燥核桃雄花为原料,在单因素试验基础上,选择料液比、提取时间、乙醇体积分数和提取温度4个因素进行雄花多酚提取工艺优化,确定超声效应下雄花多酚提取的较佳工艺条件。以超氧阴离子和羟基自由基的清除率为评价指标,对雄花多酚的抗氧化性进行研究;采用高效液相色谱对核桃雄花中多酚物质种类进行鉴定并定量分析,最后通过扫描电镜对核桃雄花超声处理前后微观结构进行表征。【结果】确定超声功率为300 W,采用多元二次回归方程模型分析得到核桃雄花多酚提取工艺参数:料液比1∶41、提取时间30 min、乙醇体积分数50%、提取温度52℃,在此条件下多酚提取率为21.32 mg/g,其中料液比、提取温度及料液比与乙醇体积分数交互作用对多酚提取率有极显著影响(P<0.01),料液比与提取时间交互作用有显著影响(P<0.05);核桃雄花多酚体外自由基清除对比试验结果表明,抗坏血酸抗氧化效果最佳,核桃雄花多酚较佳,2,6-二叔丁基对甲酚(BHT)最弱;采用高效液相色谱仪从核桃雄花中鉴定出4种单体酚,其中芦丁含量(804 μg/g)最高,绿原酸含量(181 μg/g)次之,槲皮素含量(41 μg/g)较低,没食子酸含量(18 μg/g)最低;扫描电镜分析结果表明超声处理破坏了核桃雄花表面的组织结构,利于多酚物质渗出。【结论】多元二次回归方程模型能较好的模拟和预测核桃雄花多酚提取率,雄花多酚提取率是多种影响因素共同互相作用的复杂结果;核桃雄花富含多酚类物质,具有较强抗氧化活性,可作为天然抗氧化剂材料来源加以开发利用。

     

    Abstract: 【Objective】 The surface response method was used to optimize the polyphenol extraction process of walnut male flowers, the composition and antioxidant activity were analyzed to provide technical support for the comprehensive utilization of walnut male flowers.【Method】 Dry walnut male flowers was used as the material. On the basis of single factor experiment, the solid to liquid ratio, extraction time, ethanol volume fraction, extraction temperature were selected to optimize the polyphenol extraction process of walnut male flowers, and the optimal extraction process conditions for polyphenol of walnut male flowers was determined under ultrasound effect, and the scavenging rate of superoxide anion and hydroxyl radicals were as indicators to study the antioxidant activities of walnut male flowers polyphenol. The types of polyphenol of walnut male flowers were identified and analyzed by high performance liquid chromatography. Finally, the microstructure of walnut male flowers powders before and after the extraction was characterized by scanning electron microscope.【Result】 The results showed that the ultrasound power was 300 W, and the optimization process parameters of polyphenol of walnut male flowers were obtained by multivariable quadratic regression equation model:solid to liquid ratio was 1:41, extraction time 30 min, ethanol volume fraction 50% and extraction temperature 52℃, the polyphenol yield of walnut male flowers under this condition was 21.32 mg/g, the solid to liquid ratio, extraction temperature and the interaction between solid to liquid ratio and ethanol volume fraction had extremely significant impact on the extraction rate of polyphenols(P<0.01), and the interaction between solid to liquid ratio and extraction time had a significant effect (P<0.05). Through the test of clearing the free radicals in vitro, it was found that vitamin C had the best antioxidant effect, walnut male flowers polyphenol were better, and the 2, 6-di-tert-butyl-4-methylphenol(BHT) was the weakest. Four kinds of monophenol were identified from walnut male flowers by high performance liquid chromatography, the rutinum content was the highest(804 μg/g), followed by chlorogenic acid content(181 μg/g), quercetin content was low(41 μg/g), and gallic acid content was the lowest(18 μg/g). The results of scanning electron microscope(SEM) showed that ultrasound treatment destroyed the tissue structure of walnut male flowers and facilitated the infiltration of polyphenols.【Conclusion】 The multiple quadratic regression equation model can simulate and predict the polyphenols extraction rate of walnut male flower, the polyphenols extraction rate of male flowers is a complex result of the interaction of many influencing factors. Walnut male flowers are rich in polyphenols, and has strong antioxidant activity, which can be developed and utilized as a source of natural antioxidant materials.

     

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