| Food sciences |
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| Separation of different polyphenols in blueberries and comparison of their protective activity on cellular oxidative damage |
| LIU Yixiang1, WU Yongpei1, CHEN Jun1,2, JI Baoping3* |
| (1. College of Bioengineering, Jimei University, Xiamen, Fujian 361021, China; 2. College of Chemical Engineering, Huaqiao University, Xiamen, Fujian 361021, China; 3. College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China) |
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Abstract Epidemiological studies reveal that diets rich in fruits and vegetables can reduce the incidence of chronic diseases, such as ageing, cardiovascular disease and cancer, et al. According to previous reports, oxidative stress is closely related with the occurrence and development of chronic diseases. Therefore, anti-oxidative compounds derived from plant foods are gaining more interest in ameliorating adverse health risks because of their low toxicity and few side effects. The World Health Organization has emphasized that phenolic components, especially from colorful fruits, play an important role in preventing diseases that seriously threaten people’s health, including cardiovascular diseases, diabetes, cancer, and obesity. Blueberries are highly recommended because of their abundant anthocyanins and other phenolic compounds. Furthermore, wild blueberries contain usually more polyphenol content than the cultivars. In China, the Greater Hinggan Mountains in Northeast China is the main producing area of wild blueberries, and the annual yield is about 100 to 200 thousand tons. Therefore, Chinese wild blueberries have great value of development and utilization. However, there are few literatures available about the nutritional compounds and anti-oxidative activity of the Chinese wild blueberries. In this study, contents of functional compounds in fresh wild blueberries, such as total phenols, total anthocyanins and vitamin C, were analyzed by FolinCiocalteu method, pHdifferential method and high performance liquid chromatography (HPLC) method, respectively. Moreover, in order to understand the compositions of polyphenols and the main antioxidative components in wild blueberries, blueberry polyphenols were separated and purified. After being purified sequentially by Amberlite XAD-7 column and Sephadex-LH-20 column, polyphenols from blueberries were subsequently separated into three fractions: anthocyaninrich fraction, phenolic acidrich fraction, and flavonoidrich fraction. The conditions of 2, 2’-azobis (2-amidinopropane) dihydrochloride (AAPH)induced oxidative damage of HepG-2 cells were optimized, including cellular proliferation capability, lactate dehydrogenase (LDH) level, and intracellular reactive oxygen species (ROS). Based on the in vitro model of AAPHinduced oxidative damage of HepG-2 cells, the antioxidative activity and synergistic effect of these polyphenolic fractions from blueberries were studied. The results indicated that the contents of total polyphenols and anthocyanin and vitamin C in wild blueberries were 459 mg, 277 mg and 2 721 μg per 100 g respectively. Among these three polyphenolic fractions, anthocyanins and phenolic acids were dominant and their contents in total blueberry polyphenols were 40.21% and 38.27% respectively, however, the content of flavonoidrich fraction was only 4.45%. After optimizing the conditions of AAPHinduced oxidative damage of HepG-2 cells, 0.6 mmol/L AAPH was selected at last due to the lower cytotoxicity and significantly higher intracellular ROS level under this AAPH concentration. On the aspect of antioxidative activity, the maximal clearance rates of ROS for anthocyaninrich fraction, phenolicrich fraction, and flavonoidrich fraction were 37%, 49%, 53% respectively, and additive effect was observed, but there were not synergistic effect among them. According to the above described results, Chinese wild blueberries are rich in anthocyanins and phenolic acids, and their good antioxidative activity is attributed to the high content of total polyphenols. It is concluded that wild blueberries are good antioxidative foods.
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Published: 20 July 2013
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蓝莓不同多酚物质的分离与抑制细胞氧化损伤功能的比较
以大兴安岭野生蓝莓为原料，通过Folin-Ciocalteu法、pH示差法和高效液相色谱法分析蓝莓中的总酚、总花色苷和维生素C(Vc)等功能成分的含量；采用XAD-7大孔树脂、Sephadex-LH-20葡聚糖凝胶柱层析法对蓝莓中的不同多酚组分进行分步纯化；以2,2’-偶氮二异丁基脒二盐酸（AAPH）诱导HepG-2细胞氧化损伤为评价模型，比较不同蓝莓多酚组分对抑制细胞氧化损伤的功能，并探讨组分间的协同作用。结果表明：每100 g蓝莓鲜果中含有459 mg总酚、277 mg花色苷和2 721 μg VC；在多酚类物质中，蓝莓主要以花色苷和酚酸为主，分别占总酚的40.21%和38.27%，其次为黄酮类，占4.45%；在蓝莓多酚的3个组分中，花色苷、酚酸和黄酮对HepG2细胞内活性氧自由基（reactive oxygen species，ROS）的最大清除率分别为37%、49%和53%，且组分间在清除细胞内ROS时存在加和作用，但无协同增效效果。综上所述，大兴安岭野生蓝莓富含多酚类物质，具有良好的抗氧化活性；除花色苷外，酚酸也是蓝莓中的主要抗氧化活性物质；因此，蓝莓良好的抗氧化活性与其丰富的总酚含量密切相关，这为利用大兴安岭野生蓝莓开发功能性食品提供了理论依据.
关键词：
蓝莓,
多酚,
抗氧化,
协同作用
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