柱层析提取黑果腺肋花楸果多种活性成分及体外活性研究

Study on Column Chromatographic Extraction of Active Components from Aronia Melanocarpa and Activity in Vitro

探索一种从黑果腺肋花楸果中分离总黄酮、总皂苷、花色苷和多糖的方法,并对各组分的抗氧化活性、抑制酪氨酸酶活性和抑制α-葡萄糖苷酶活性的能力进行考察。以目标组分的质量为评价指标,确定了总黄酮、总皂苷、花色苷和多糖的最佳提取溶剂分别为无水乙醇、60%乙醇、60%乙醇和水,吸涨率分别为1.5、2.5、2.5、2.5 mL/g,浸泡平衡时间分别为2、2、2、1 h;用半数浓度(IC_(50)值)作为指标来评价各组分的不同体外活性。结果表明循环提取第1轮总黄酮、总皂苷、花色苷和多糖的提取率分别是76.09%、84.38%、73.75%、72.23%,第4轮开始各组分的提取率均达到90%以上,分别得总黄酮、总皂苷、花色苷和多糖的粗产品,含量分别为8.93±0.16、70.39±0.26、1.51±0.02和695.46±1.39 mg/g,经纯化处理后总黄酮、总皂苷、花色苷和多糖干浸膏中的各组分的含量分别为30.29±0.24、252.78±0.73、3.66±0.24和884.62±2.03 mg/g,体外活性实验表明各组分均具有一定的抗氧化活性、酪氨酸酶活性抑制和α-葡萄糖苷酶活性抑制的能力。柱层析循环法联合提取可以同时高效地分离多种活性成分,提取所得的各组分均具有不同的体外活性,实验结果为黑果腺肋花楸果的食品开发与利用提供实验依据和技术支持。

A method was developed for the separation of total flavonoids,total saponins,anthocyanins and polysaccharides from Aronia melanocarpa,and their antioxidant activity,inhibition of tyrosinase activity and inhibition ofα-glucosidase activity were investigated also.Using quality of target components as evaluation index,the optimal extraction parameter of total flavonoids,total saponins,anthocyanins and polysaccharides were ascertained as follows,the most suitable solvents as anhydrous ethanol,60%ethanol,60%ethanol and water,the absorption rate as 1.5,2.5,2.5,2.5 mL/g,and the soaking equilibrium time as 2,2,2,1 h,respectively.IC_(50) value was used as index to evaluate the activity difference in vitro between each component.The results showed during the first round of cyclic extraction the extraction rates of total flavonoids,total saponins,anthocyanins and polysaccharides were 76.09%,84.38%,73.75%and 72.23%,which that of all components reached more than 90%during the fourth round.The concentration of total flavonoids,total saponins,anthocyanins and polysaccharides in crude products were 8.93±0.16,70.39±0.26,1.51±0.02 and 695.46±1.39 mg/g,respectively,and they were purified to the concentration of 30.29±0.24,252.78±0.73,3.66±0.24 and 884.62±2.03 mg/g,respectively.Experiments on activity in vitro indicated that all the components had certain antioxidant activity,tyrosinase activity inhibition andα-glucosidase activity inhibition ability.Column chromatographic method combined with cyclic extraction could separate efficiently multiple active components which all possess different activities in vitro.The research results provided experimental basisand technical support for Aronia melanocarpa in food development and utilization field.