Ralls苹果多酚分离及抗氧化与脂肪代谢调节作用研究
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摘要
苹果多酚作为天然的抗氧化物可以抑制或阻断氧化过程、平衡自由基,所以引起研究者的广泛关注。本文以Ralls苹果为研究对象,采用研磨珠辅助超声波法确定了Ralls苹果多酚及其抗氧化物质的最佳提取工艺,采用大孔树脂吸附分离技术制备了苹果多酚,对其抗氧化和脂肪代谢调节功能进行了研究,旨在为苹果多酚的进一步开发利用提供参考。具体研究结果如下：
(1)单因素研究结果表明研磨珠萃取法,可以有效提取Ralls苹果多酚。
(2)响应面法确定了Ralls苹果总酚最佳提取工艺为：超声时间22.62 min,超声温度50.38℃,超声功率243.85 W,研磨珠加载量(珠：料g/g)1.25:1；Ralls苹果总黄酮最佳提取工艺为：超声时间22.75 min,超声温度51.52℃,超声功率229.35 W,研磨珠加载量(珠：料g/g)1.21:1；Ralls苹果原花青素最佳提取工艺为：超声时间21.17 min,超声温度51.09℃,超声功率249.83 W,研磨珠加载量(珠：料g/g)1.22:1；苹果多酚类抗氧化物质的最佳提取工艺为：超声时间23.75 min,超声温度：50.81℃,超声功率244.8W,研磨珠加载量(珠：料g/g)1.21:1。
(3)通过静态吸附、解析实验确定分离Ralls苹果多酚的最佳固定相为X-5型大孔树脂,制得Ralls苹果多酚纯度可达70%。
(4)对Ralls苹果多酚进行体外抗氧化活性实验,结果表明苹果多酚具有很强的总抗氧化能力和还原能力,并且可以有效的清除羟自由基、超氧自由基、DPPH自由基和过氧化氢自由基。
(5)采用建立高脂动物模型的方法,研究苹果多酚对小鼠脂肪代谢的调节作用,结果表明,苹果多酚提取物和苹果多酚纯化物均能明显降低小鼠血中甘油三酯、总胆固醇、低密度脂蛋白(LDL-C)和MDA含量以及动脉粥样硬化指数(AI),对小鼠血清中HDL-C有明显的升高作用,能有效提高小鼠体内超氧化物歧化酶、谷胱甘肽过氧化物酶、脂蛋白脂酶和肝脂酶的活性,从而具有降脂、调节脂肪代谢的功能。
(6)对Ralls苹果与其他品种苹果的成分与功能比较结果表明,Ralls苹果的总酚含量要高于其他六个品种苹果,而总黄酮含量和原花青素含量则分别次于铃铛果和黄太平苹果,位居第二。Ralls苹果的总抗氧化能力仅次于黄太平苹果,位居第二；清除羟自由基的能力仅次于77-6,也位居第二。
As natural antioxidant, apple polyphenols can restrain or interdicte oxidation process and balance the free radicles, so many reserachers had gave broad attention on it. The Ralls apple was selected as experiment material in this paper. The best extraction technics for apple polyphenols and it's antioxidant substance were studied through ultrasonic method with the assistant of grind balls. The apple polyphenols was prepared through the adsorption and separation technic using macroporous resin, and the function of antioxidation and regulation fat metabolization were also studied in order to provied reference for more developing and using of the apple polyphenols. The results showed as follows:
(1) The result of single factor experiment showed that extraction method with grind balls could extract Ralls apple polyphenols very efficiently.
(2) The best extraction technics for Ralls apple polyphenols and it's antioxidant substance were obtained by surface response method. The best extraction technics for Ralls apple total polyphenols is that:the ultrasonic time is 22.62 min, the ultrasonic temperature is 50.38℃, the ultrasonic power is 243.85 W, the ratio of balls to material is 1.25:1. The best extraction technics for Ralls apple total flavone is that:the ultrasonic time is 22.75 min, the ultrasonic temperature is 51.52℃, the ultrasonic power is 229.35 W, the ratio of balls to material is 1.21:1. The best extraction technics for Ralls apple anthocyanin is that:the ultrasonic time is 21.17 min, the ultrasonic temperature is 51.09℃, the ultrasonic power is 249.83 W, the ratio of balls to material is 1.22:1. The best extraction technics for Ralls apple antioxidant substance is that:the ultrasonic time is 23.75 min, the ultrasonic temperature is 50.81℃, the ultrasonic power is 244.8 W, the ratio of balls to material is 1.21:1.
(3) The stationary phases for extration of Ralls apple polyphenols were established by static adsorption and analytical experiment. The best stationary phase was X-5, and the pure of Ralls apple polyphenols extracted by this stationary phase could reach seventy percent.
(4) The results of extracorporeal antioxidation experiment showed that apple polyphenols had very strong total antioxidant ability and reduction ability, and it could clean up hydroxyl free radical, superoxide anion radical, DPPH free radical and hydrogen peroxide effectively.
(5) The method of establishing high fat animal model was adopted to research the effect of apple polyphenols on lipid metobolism in mice. The results showed that apple polyphenols could obviously reduce the content of TQ TC, low density lipoprotein cholesterol(LDL-C) and MDA, and atherosclerosisdisease index(AI). It could also improve HDL-C and the activity of SOD、GSH-PX、HL and LPL in their blood serum. The apple polyphenols have the functions in reducing blood lipid by adjusting the lipid metobolism.
(6) The componet and function of Rails apple were compared with other apple varieties, and the results showed that the total polyphenols content of Rails apple was higher than other six apple varities, but the total flavone and anthocyanin content of Rails apple ranked the second place in all the seven varieties, it was lower than Small Bell apple and "Hangtaiping" apple respectively. The total antioxidant ability of Rails apple ranked the second plance, it was lower than "Hangtaiping" apple, and the hydroxyl free radical cleaning up ability of Rails apple ranked also the second plance, it was lower than "77-6" apple.
(1)单因素研究结果表明研磨珠萃取法,可以有效提取Ralls苹果多酚。
(2)响应面法确定了Ralls苹果总酚最佳提取工艺为：超声时间22.62 min,超声温度50.38℃,超声功率243.85 W,研磨珠加载量(珠：料g/g)1.25:1；Ralls苹果总黄酮最佳提取工艺为：超声时间22.75 min,超声温度51.52℃,超声功率229.35 W,研磨珠加载量(珠：料g/g)1.21:1；Ralls苹果原花青素最佳提取工艺为：超声时间21.17 min,超声温度51.09℃,超声功率249.83 W,研磨珠加载量(珠：料g/g)1.22:1；苹果多酚类抗氧化物质的最佳提取工艺为：超声时间23.75 min,超声温度：50.81℃,超声功率244.8W,研磨珠加载量(珠：料g/g)1.21:1。
(3)通过静态吸附、解析实验确定分离Ralls苹果多酚的最佳固定相为X-5型大孔树脂,制得Ralls苹果多酚纯度可达70%。
(4)对Ralls苹果多酚进行体外抗氧化活性实验,结果表明苹果多酚具有很强的总抗氧化能力和还原能力,并且可以有效的清除羟自由基、超氧自由基、DPPH自由基和过氧化氢自由基。
(5)采用建立高脂动物模型的方法,研究苹果多酚对小鼠脂肪代谢的调节作用,结果表明,苹果多酚提取物和苹果多酚纯化物均能明显降低小鼠血中甘油三酯、总胆固醇、低密度脂蛋白(LDL-C)和MDA含量以及动脉粥样硬化指数(AI),对小鼠血清中HDL-C有明显的升高作用,能有效提高小鼠体内超氧化物歧化酶、谷胱甘肽过氧化物酶、脂蛋白脂酶和肝脂酶的活性,从而具有降脂、调节脂肪代谢的功能。
(6)对Ralls苹果与其他品种苹果的成分与功能比较结果表明,Ralls苹果的总酚含量要高于其他六个品种苹果,而总黄酮含量和原花青素含量则分别次于铃铛果和黄太平苹果,位居第二。Ralls苹果的总抗氧化能力仅次于黄太平苹果,位居第二；清除羟自由基的能力仅次于77-6,也位居第二。
As natural antioxidant, apple polyphenols can restrain or interdicte oxidation process and balance the free radicles, so many reserachers had gave broad attention on it. The Ralls apple was selected as experiment material in this paper. The best extraction technics for apple polyphenols and it's antioxidant substance were studied through ultrasonic method with the assistant of grind balls. The apple polyphenols was prepared through the adsorption and separation technic using macroporous resin, and the function of antioxidation and regulation fat metabolization were also studied in order to provied reference for more developing and using of the apple polyphenols. The results showed as follows:
(1) The result of single factor experiment showed that extraction method with grind balls could extract Ralls apple polyphenols very efficiently.
(2) The best extraction technics for Ralls apple polyphenols and it's antioxidant substance were obtained by surface response method. The best extraction technics for Ralls apple total polyphenols is that:the ultrasonic time is 22.62 min, the ultrasonic temperature is 50.38℃, the ultrasonic power is 243.85 W, the ratio of balls to material is 1.25:1. The best extraction technics for Ralls apple total flavone is that:the ultrasonic time is 22.75 min, the ultrasonic temperature is 51.52℃, the ultrasonic power is 229.35 W, the ratio of balls to material is 1.21:1. The best extraction technics for Ralls apple anthocyanin is that:the ultrasonic time is 21.17 min, the ultrasonic temperature is 51.09℃, the ultrasonic power is 249.83 W, the ratio of balls to material is 1.22:1. The best extraction technics for Ralls apple antioxidant substance is that:the ultrasonic time is 23.75 min, the ultrasonic temperature is 50.81℃, the ultrasonic power is 244.8 W, the ratio of balls to material is 1.21:1.
(3) The stationary phases for extration of Ralls apple polyphenols were established by static adsorption and analytical experiment. The best stationary phase was X-5, and the pure of Ralls apple polyphenols extracted by this stationary phase could reach seventy percent.
(4) The results of extracorporeal antioxidation experiment showed that apple polyphenols had very strong total antioxidant ability and reduction ability, and it could clean up hydroxyl free radical, superoxide anion radical, DPPH free radical and hydrogen peroxide effectively.
(5) The method of establishing high fat animal model was adopted to research the effect of apple polyphenols on lipid metobolism in mice. The results showed that apple polyphenols could obviously reduce the content of TQ TC, low density lipoprotein cholesterol(LDL-C) and MDA, and atherosclerosisdisease index(AI). It could also improve HDL-C and the activity of SOD、GSH-PX、HL and LPL in their blood serum. The apple polyphenols have the functions in reducing blood lipid by adjusting the lipid metobolism.
(6) The componet and function of Rails apple were compared with other apple varieties, and the results showed that the total polyphenols content of Rails apple was higher than other six apple varities, but the total flavone and anthocyanin content of Rails apple ranked the second place in all the seven varieties, it was lower than Small Bell apple and "Hangtaiping" apple respectively. The total antioxidant ability of Rails apple ranked the second plance, it was lower than "Hangtaiping" apple, and the hydroxyl free radical cleaning up ability of Rails apple ranked also the second plance, it was lower than "77-6" apple.
引文
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