虎杖根茎和桂花果肉的抗氧化活性初探
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摘要
从天然资源中寻找具有抗氧化活性的有效成分是发现天然抗氧化剂的重要途径之一,传统或民间药用植物是其中一个非常重要的来源。本文运用一系列的抗氧化性能测试方法:包括DPPH自由基清除能力测定、羟基自由基清除能力测定、ABTS自由基清除能力测定、脂质抗氧化能力测定、还原能力、总抗氧化能力、总酚含量测定、总黄酮含量测定等方法对虎杖和桂花果肉的抗氧化活性进行了初探。
第一部分:虎杖抗氧化活性初探
中药虎杖为寥科植物虎杖(Polygonum cuspidatum Siebet Zucc)的根茎,它具有广泛的生物活性,临床上常用于治疗动脉硬化、水火烫伤、利尿、跌打损伤及化脓性皮炎等。本研究对虎杖乙醇提取物、总蒽醌及其中的三种不同酸性成分进行了抗氧化活性初筛,结果表明虎杖总蒽醌和其中的强酸成分具有很强的抗氧化活性,活性高于合成抗氧化剂BHT及已经投入市场的天然抗氧化剂白藜芦醇。随后分离出四种虎杖总蒽醌中的常见成分,大黄酸、大黄素、大黄酚和大黄素甲醚,并对其抗氧化活性进行了测定,发现其中仅有大黄酸的活性与白藜芦醇接近,其它均弱于白藜芦醇和BHT,因此,推断虎杖总蒽醌及其中的强酸成分的高活性是各单体化合物协同作用的结果。进而将总蒽醌和强酸成分具体运用到食品中,实验结果显示,虎杖总蒽醌及其强酸成分在食品中的抗氧化活性明显高于合成抗氧化剂BHT。因此,二者有望开发成天然抗氧化剂。
第二部分:桂花果肉抗氧化活性初探
桂花(O. fragrans)系木犀科(Oleaceae)木犀属(Osmanthus)植物,其花、果、根均可入药,民间用作止痛剂,治疗肝胃气痛等。由于桂花果仅有少量用于繁殖桂花树,大量的果实散落下来,不仅浪费资源,同时对环境也造成了很大的污染。本研究通过活性引导法对桂花果肉的抗氧化活性部位和活性成分进行了初探。活性筛选结果显示,桂花果肉乙醇提取物的甲醇可溶部分(MSF)为活性部位。并对其进行了急性毒理试验,结果显示,其对小鼠的最大耐受量均大于34.28g/kg,即该部位无毒。进而对MSF进行了分离,从中首次分离出了两个单体化合物,分别为oleoside dimethyl ester和红景天甙。其中红景天甙的活性明显高于所有粗提物及合成抗氧化剂BHT,因此可确定红景天甙是桂花果肉中的主要抗氧化活性化合物之一。本研究为桂花果肉的开发和利用提供了理论依据。
Searching for antioxidant active component from natural origins remains is one of the most important approaches in discovering natural antioxidants, and traditional medicinal plants or folk herbs have proved to be very important sources. In this thesis, antioxidant capability of Polygonum cuspidatum and Osmanthus fragrans pulp was compared and estimated by several established biochemical assays in vitro: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals scavenging assay, 2,2’-azino-bis (3- ethylbenzothiazoline- 6-sulfonic acid) diamonium salt (ABTS) free radicals scavenging assay, reducing power assay which assess electron donation capacity and phosphomolybdenum assay which investgate total antioxidant capacity, Total phenolics content and Total flavone content.
Part 1: Study on antioxidant activity of Polygonum cuspidatum
Polygonum cuspidatum S. et Z. belongs to the Polygonaceae perennial herb group widely distributed in China. It was traditionally used in China and Japan as a folk medicine for the treatment of atherosclerosis and for other therapeutic purposes such as scalds, burns, laxation, a diuretic, and for the treatment of suppurative dermatitis in the oriental medicine. In this thesis, antioxidant activities of Polygonum cuspidatum ethanol extract (EE), total anthraquinones (TA) and three different acid parts were compared. It is showed that TA and strong acid part (SAP) exhibited excellent antioxidant activity than that of BHT and resveratrol. Four activities compounds were isolated from TA. There are rhein, emodin, chrysophanol and physcion. But antioxidant activities of these compounds were weaker than BHT and resveratrol expect rhein which was near to resveratrol. Therefore, the strong antioxidant activity of TA and SPA was the cooperation of all the single compounds. Moreover, TA and SPA were appended into food as a food additive. The research investigated that antioxidant activtity of TA and SPA was stronger than BHT. Hence, they are expected to be empoldered as natural antioxidants.
Part 2: Study on antioxidant activity of Osmanthus fragrans pulp
The flowers, fruits and roots of O. fragrans, a species of the Oleaceae family, have been widely used in the TCM or folklore medicine to treat liver and stomach pains. Only minorty O. fragrans seeds are keeping as the seed cultivating in the becoming year. The majority of the seeds are left and regards as a waste material. And it may cause the environmental problems. In this thesis, antioxidant activities of O. fragrans was estimated and filtrated. The filtration result indicated that methanol soluble fraction (MSF) extracted from crude ethanol extract was the activity component. Furthermore, acute toxicity study in rats of provided that MSF was non-toxic (MTD (Maximum Tolerated Dose) in rat’s > 34.28 g/kg). Two known compounds were isolated from the O. fragran for the first time. The structures of these compounds were identified to be Oleoside dimethyl ester (1) and salidroside (2). And the antioxidant properties of two compounds also evaluated through these established methods. Results from the antioxidant properties evaluation show that salidroside exerted powerful antioxidant activities in all test system. Therefor, salidroside had great potential as natural antioxidant. This study provides a basical date for value exploitation and utilization of O. fragran pulp.
第一部分:虎杖抗氧化活性初探
中药虎杖为寥科植物虎杖(Polygonum cuspidatum Siebet Zucc)的根茎,它具有广泛的生物活性,临床上常用于治疗动脉硬化、水火烫伤、利尿、跌打损伤及化脓性皮炎等。本研究对虎杖乙醇提取物、总蒽醌及其中的三种不同酸性成分进行了抗氧化活性初筛,结果表明虎杖总蒽醌和其中的强酸成分具有很强的抗氧化活性,活性高于合成抗氧化剂BHT及已经投入市场的天然抗氧化剂白藜芦醇。随后分离出四种虎杖总蒽醌中的常见成分,大黄酸、大黄素、大黄酚和大黄素甲醚,并对其抗氧化活性进行了测定,发现其中仅有大黄酸的活性与白藜芦醇接近,其它均弱于白藜芦醇和BHT,因此,推断虎杖总蒽醌及其中的强酸成分的高活性是各单体化合物协同作用的结果。进而将总蒽醌和强酸成分具体运用到食品中,实验结果显示,虎杖总蒽醌及其强酸成分在食品中的抗氧化活性明显高于合成抗氧化剂BHT。因此,二者有望开发成天然抗氧化剂。
第二部分:桂花果肉抗氧化活性初探
桂花(O. fragrans)系木犀科(Oleaceae)木犀属(Osmanthus)植物,其花、果、根均可入药,民间用作止痛剂,治疗肝胃气痛等。由于桂花果仅有少量用于繁殖桂花树,大量的果实散落下来,不仅浪费资源,同时对环境也造成了很大的污染。本研究通过活性引导法对桂花果肉的抗氧化活性部位和活性成分进行了初探。活性筛选结果显示,桂花果肉乙醇提取物的甲醇可溶部分(MSF)为活性部位。并对其进行了急性毒理试验,结果显示,其对小鼠的最大耐受量均大于34.28g/kg,即该部位无毒。进而对MSF进行了分离,从中首次分离出了两个单体化合物,分别为oleoside dimethyl ester和红景天甙。其中红景天甙的活性明显高于所有粗提物及合成抗氧化剂BHT,因此可确定红景天甙是桂花果肉中的主要抗氧化活性化合物之一。本研究为桂花果肉的开发和利用提供了理论依据。
Searching for antioxidant active component from natural origins remains is one of the most important approaches in discovering natural antioxidants, and traditional medicinal plants or folk herbs have proved to be very important sources. In this thesis, antioxidant capability of Polygonum cuspidatum and Osmanthus fragrans pulp was compared and estimated by several established biochemical assays in vitro: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals scavenging assay, 2,2’-azino-bis (3- ethylbenzothiazoline- 6-sulfonic acid) diamonium salt (ABTS) free radicals scavenging assay, reducing power assay which assess electron donation capacity and phosphomolybdenum assay which investgate total antioxidant capacity, Total phenolics content and Total flavone content.
Part 1: Study on antioxidant activity of Polygonum cuspidatum
Polygonum cuspidatum S. et Z. belongs to the Polygonaceae perennial herb group widely distributed in China. It was traditionally used in China and Japan as a folk medicine for the treatment of atherosclerosis and for other therapeutic purposes such as scalds, burns, laxation, a diuretic, and for the treatment of suppurative dermatitis in the oriental medicine. In this thesis, antioxidant activities of Polygonum cuspidatum ethanol extract (EE), total anthraquinones (TA) and three different acid parts were compared. It is showed that TA and strong acid part (SAP) exhibited excellent antioxidant activity than that of BHT and resveratrol. Four activities compounds were isolated from TA. There are rhein, emodin, chrysophanol and physcion. But antioxidant activities of these compounds were weaker than BHT and resveratrol expect rhein which was near to resveratrol. Therefore, the strong antioxidant activity of TA and SPA was the cooperation of all the single compounds. Moreover, TA and SPA were appended into food as a food additive. The research investigated that antioxidant activtity of TA and SPA was stronger than BHT. Hence, they are expected to be empoldered as natural antioxidants.
Part 2: Study on antioxidant activity of Osmanthus fragrans pulp
The flowers, fruits and roots of O. fragrans, a species of the Oleaceae family, have been widely used in the TCM or folklore medicine to treat liver and stomach pains. Only minorty O. fragrans seeds are keeping as the seed cultivating in the becoming year. The majority of the seeds are left and regards as a waste material. And it may cause the environmental problems. In this thesis, antioxidant activities of O. fragrans was estimated and filtrated. The filtration result indicated that methanol soluble fraction (MSF) extracted from crude ethanol extract was the activity component. Furthermore, acute toxicity study in rats of provided that MSF was non-toxic (MTD (Maximum Tolerated Dose) in rat’s > 34.28 g/kg). Two known compounds were isolated from the O. fragran for the first time. The structures of these compounds were identified to be Oleoside dimethyl ester (1) and salidroside (2). And the antioxidant properties of two compounds also evaluated through these established methods. Results from the antioxidant properties evaluation show that salidroside exerted powerful antioxidant activities in all test system. Therefor, salidroside had great potential as natural antioxidant. This study provides a basical date for value exploitation and utilization of O. fragran pulp.
引文
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