刘皓涵, 钟迪颖, 张润光, 王国良, 张有林. 欧李多酚提取纯化及抗氧化性研究[J]. 农业工程学报, 2020, 36(22): 324-332. DOI: 10.11975/j.issn.1002-6819.2020.22.036
    引用本文: 刘皓涵, 钟迪颖, 张润光, 王国良, 张有林. 欧李多酚提取纯化及抗氧化性研究[J]. 农业工程学报, 2020, 36(22): 324-332. DOI: 10.11975/j.issn.1002-6819.2020.22.036
    Liu Haohan, Zhong Diying, Zhang Runguang, Wang Guoliang, Zhang Youlin. Extraction and purification of polyphenols and determination of antioxidant activity[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2020, 36(22): 324-332. DOI: 10.11975/j.issn.1002-6819.2020.22.036
    Citation: Liu Haohan, Zhong Diying, Zhang Runguang, Wang Guoliang, Zhang Youlin. Extraction and purification of polyphenols and determination of antioxidant activity[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2020, 36(22): 324-332. DOI: 10.11975/j.issn.1002-6819.2020.22.036

    欧李多酚提取纯化及抗氧化性研究

    Extraction and purification of polyphenols and determination of antioxidant activity

    • 摘要: 欧李富含钙素和多酚类物质,具有提高人体免疫力等保健功能,近年来在中国得到了大力发展。欧李多酚提取、纯化及其抗氧化功能,为欧李开发利用提供理论依据。该研究采用单因素及响应面优化试验研究了欧李多酚超声辅助酶解提取工艺,采用大孔树脂纯化多酚粗提物,利用高效液相色谱法测定酚类物质的种类,利用分光光度法测定欧李多酚类物质总还原力和对自由基·OH、DPPH·、ABTS·的清除作用。试验得出欧李多酚提取最优工艺参数为:超声波功率105 W、酶解温度50 ℃、酶解时间80 min、液料比30∶1 mL/g,多酚提取量为42.63 mg/g。经大孔树脂纯化后纯化物中欧李总多酚含量为73.42 mg/g,从欧李果实中检测出6种多酚类物质,其含量顺序为绿原酸>没食子儿茶素没食子酸酯>咖啡因>对羟基苯甲酸>原儿茶酸>没食子酸。欧李多酚的总还原力与维生素C相当,对·OH、DPPH·、ABTS·等自由基清除作用显著。

       

      Abstract: Abstract: Cerasus humilis is naturally rich in a variety of vital vitamins and minerals, such as calcium and polyphenols, providing great benefits for health care functions, particularly on improving human immunity. Polyphenols are significant natural antioxidant substances for various functions, including softening blood vessels, promoting digestion, lowering blood lipids, diuresis, enhancing immunity, preventing arteriosclerosis and thrombus formation, even to inhibit the growth of bacteria and cancer. Specifically, an emphasis is put effect on inhibiting the proliferation of human liver cancer (HepG2), colon cancer (HCT116), and gastric cancer (BGC823). However, only a few reports focused on the natural active substances that contained in plum fruit. In this study, a novel extraction and purification process of plum polyphenols was proposed to determine the types and contents of polyphenols, and thereby to explore the in vitro effects of polyphenols. The processing parameters were optimized in the extraction and purification of polyphenols, together with the antioxidant activity of plum cerasus humilis. A single factor and response surface optimization tests were used to investigate the ultrasonic assisted enzymatic extraction process of polyphenol of Cerasus humilis. Macroporous resin was also selected to purify the crude extract of polyphenols. Moreover, high solution liquid chromatography was used to identify the types of phenolic substances, and the spectrophotometry was used to determine the total reducing power of polyphenol in cerasus humilises, and the scavenging effect of free radicals ·OH, DPPH·, ABTS·. The results showed that the optimum extraction parameters of polyphenol of cerasus humilis were as follows: The ultrasonic power was 105 W, temperature of enzymatic hydrolysis was 50 ℃, enzymatic hydrolysis time was 80 min, the ratio of liquid to material was 30:1 (mL/g), and the extraction amount of polyphenols was 42.63 mg/g. After the purification with macroporous resin, the total polyphenol content of cerasus humilises was 73.42 mg/g. All the 6 polyphenols were detected in the plum fruit of cerasus humilises, where their contents were ranked in order, chlorogenic acid > epigallocatechin gallate> caffeine > p-hydroxybenzoic acid >protocatechuic acid gallic > gallic acid. The total reducing power of polyphenol in cerasus humilises was not different from that of Vitamin C, indicating a significant scavenging effect on free radicals, such as ·OH, DPPH·, ABTS·. In the case of the same mass concentration, the reduction ability of purified product, crude extract, and Vc of the cerasus humilis were as follows: Vitamin C (IC50=35.05μg/mL) > the cerasus humilis polyphenols (IC50 =32.42 μg/mL) > the crude extraction of Oleum Phenol (IC50=30.13 μg/mL). The reduction ability of purified cerasus humilis was not significantly different from that of vitamin C (P>0.05), but there was totally difference from the crude extract (P<0.05). The purification process can be used to remove some impurities without affecting the activity of the main components, indicating that it can improve the total reduction capacity. Cerasus humilis showed the good scavenging ability on DPPH·, ABTS·, and ·OH radicals. The purified product of cerasus humilis demonstrated an IC50 value of 72.65 μg/mL for the removal of ·OH, and an IC50 value of 110.12 μg/mL for the removal of DPPH·. The IC50 value for the removal of ABTS· was 55.45 μg/mL, and the total reducing power EC50 value was 32.42 μg/mL, indicating equivalent to the effect of Vc. The IC50 value for the crude extract of Prunella vulgaris polyphenols was 66.92 μg/mL for removing ·OH, the IC50 value for removing DPPH· was 98.42 μg/mL, the IC50 value for removing ABTS· was 49.87 μg/mL, and the total reducing power EC50 value was 30.13 μg/mL. The finding can provide a sound theoretical basis and technical support for the development and utilization of cerasus humilis in the production of plum fruits.

       

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