NWIPB OpenIR
唐古特白刺(Nitraria tangutorum Bobr.)天然色素的研究
Alternative TitleStudy on Natural Pigment from Nitraria tangutorum Bobr.
丁晨旭
Subtype博士
Thesis Advisor索有瑞
2008-06-10
Degree Grantor中国科学院西北高原生物研究所
Place of Conferral西北高原生物研究所
Keyword唐古特白刺 红色素 花色苷 自由基 活性氧 提取 大孔树脂 纯化 稳定性 矿质元素 抗氧化 降血脂
Abstract近20年来,活性氧和自由基研究成为现代生命科学的前沿和热点,评价和筛选具有强抗氧化活性的天然资源己成为生物学、医学和食品科学研究的新趋势。活性氧可对生物体产生一系列的有害作用,例如进攻组织内的多不饱和脂肪酸,蛋白质和核酸等,造成氧化损伤,而这种氧化损伤与很多衰老疾病如心脏病、神经系统紊乱和癌症等的病因有关。 花色苷是一种在植物中普遍存在的酚类物质,是很多植物叶、花和果实呈现鲜艳颜色的原因。它具有抑制活性氧的生成和清除活性氧的生理活性,花色苷的抗氧化性源于其结构中的侧环酚羟基。 本学位论文工作对白刺果实中的花色苷及其活性进行了较为全面、系统和深入的研究,取得了以下重要研究成果和结论。 1、通过正交实验确定了白刺果实花色苷粗产品最佳提取条件为:温度:85℃;盐酸溶液浓度:1%;60%乙醇溶液;1:40的料液比;提取时间:40min;次数:2次。 2、通过静态吸附解析实验,确定了利用大孔吸附树脂初步纯化唐古特白刺果实花色苷的条件,并根据所得条件做了扩大实验。结果表明:X-5树脂的吸附能力最好,吸附率为76.85%,pH在1-3范围内对吸附效果无影响;较高的乙醇浓度和pH值有利于色素的解析,解析液以80%酸化乙醇(含1%HCl)效果最佳,解析时间为40min;扩大实验得到紫红色色素粉末,得率为1.24%,水溶性好,不含糖类、蛋白质等水溶性杂质。 3、白刺果实花色苷的稳定性研究表明:该花色苷在弱光和较低的温度下稳定性较好,对大部分金属离子较稳定,碳水化合物对花色苷的稳定性影响不大,H2O2、Na2SO3对花色苷的影响较大。 4、对唐古特白刺色素的矿物质元素进行了分析。结果表明,在色素中Na、K、Ca含量高,这与白刺生长在盐碱地有关;Fe、Zn、Cu、Mn、Se等必需微量元素含量丰富;有害元素含量低。 5、对白刺果实花色苷进行光谱和色谱两方面的研究,通过观察纯化的花色苷衍生物的颜色、加入5%三氯化铝甲醇(W:V)溶液后最大吸收波长是否发生变化、A440/Amax等光谱性质和对其完全酸水解、受控酸水解以及层析Rf值等色谱数据确定了其糖苷配基的类型以及糖基的种类和糖苷的连接方式,确定了白刺果实花色苷的主要成分为:矢车菊-3-葡萄糖苷。 6、探讨了唐古特白刺果实花色苷体外清除自由基活性,研究了其对超氧阴离子(O2-•)体系、羟基自由基(•OH)体系和二苯代苦味酰基自由基(DPPH•)体系的清除效果,并与维生素C进行了比较。结果表明,该花色苷对O2-•、•OH、DPPH• 均具有清除作用,尤其对•OH和DPPH• 清除效果相对较好,优于Vc,且与浓度呈量效关系,对O2-•的清除效果低于Vc。唐古特白刺红色素是一种较好的天然自由基清除剂 7、通过动物试验,发现白刺果实花色苷既可有效降低高脂血症大鼠的血脂水平,改善血脂代谢,减少动脉粥样硬化的危险性,又可有效提高大鼠体内的抗氧化水平,增强抗氧化酶活性,减少过氧化脂质含量。主要表现为:显著降低总甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和载脂蛋白B(ApoB)及动脉硬化指数TC-HDL-C/HDL-C和LDL-C/HDL-C(P<0.05),显著提高高密度脂蛋白胆固醇(HDL-C)和载脂蛋白A(ApoA)水平(P<0.05);此外,还显著提高大鼠血清和肝脏的总抗氧化能力(TAC)和超氧化物歧化酶(SOD)活性,降低脂质过氧化物丙二醛(MDA)含量,并呈现显著的剂量依赖性。高血脂症大鼠的血脂水平和动脉硬化指数与其氧化-抗氧化水平密切相关,二者互相促进。 8、提出并证明白刺果实花色苷降血脂和抗动脉粥样硬化等预防心血管疾病的机理主要在于其花色苷类化合物的抗氧化作用:白刺果实花色苷类化合物抗氧化的主要机制在于其酚羟基提供氢原子;阻止体内脂肪的氧化和捕捉体内活性自由基;形成酚醌平衡的反应特性。
Other AbstractIn the last 20 years, active oxygen and free radical have become the hot topics of modern life science, and appraisal and selection of natural resources with strong antioxidative activities have become the new trends of biology, medical science and food science and technology. Reactive oxygen species (ROS) are very damaging to organisms, since they can attack polyunsatural fatty acids (PUFA), proteins and nuclear acid in tissues to induce oxidation. This oxidative damage is considered to play a causative role in aging and degenerative diseases associated with it, such as heart disease, cognitive dysfunction and cancer. Anthocyanins are polyphenols widely distributed in plants, and they contribute to the brilliant color of blue, red, scarlet in leaves, flowers and fruits. It is reported that they have potentials inhibiting the formation of ROS and scavenging ROS, and the antioxidant ability due to the hydroxyl groups substituted in the aromatic B-ring. In this research, anthocyanins and its activity of Nitraria tangutorum Bobr. fruit have been studied widely、systematically and deeply. And we get some results and conclusions as followed. 1. The optimum extracting conditions of crude anthocyanins from Nitraria tangutorun Bobr. fruit were established by orthogonal experiment. The results showed that, the best extraction effect was under the conditions of 85℃, 1%Hydrochloric acid-60%ethanol, 1:40 the ratio, extracting 2 times and 40min for each times. 2. The purification conditions of anthocyanins from Nitraria tangutorum Bobr. fruit using macroporous resin were confirmed by adsorbing and desorption experiment and enlarged experiment was studied. The results showed that, X-5 resin had more strong adsorption capacity, the adsorptive rate of pigment reached 76.85%, pH 1-3 had no influence on adsorbing effect; Higher concentration of ethanol and acidity had a higher desorption ability for red pigment, 80% ethanol contained 1% Hydrochloric acid had the best desorption effect and the time of desorption were 40 minutes; We got powder of anthocyanin through enlarged experiment and the rate of anthocyanin was 1.24%, This powder had good water-solubility and did not contain some impurities such as glucide, protein. 3. Studies indicated that anthocyanins were stable in weak light, low temperature and most metal ions. Carbohydrate had few influence on anthocyanins, but Na2SO3 and H2O2 were opposite. 4. The conments of mineral elements in pigment of Nitraria tangutorun Bobr. fruit were analysized. The results show that the conments of Na, K, Ca are very abundant. It is related with Nitraria tangutorun Bobr. growing in saline or alkaline soil. The conments of essential trace elements Fe, Zn, Cu, Mn, Se are also abundant. And the conments of hazard elements are poor. 5. By the spectrum and chromatography analysis, the anthocyanins derivates spectrum properties were investigated, through study the colour, the changes of maximum absorption after adding 5%AlCl3-methanol(W:V)and A440/Amax of purified anthocyanins; the type of aglycones, sugurs and indican connect fashions were determined by complete acid hydrosis and controlled acid hydrosis. At last, the conclusion were: the main anthocyanin structure in Nitraria tangutorun Bobr. pigments is cyanidin-3-glucoside. 6. To investigate the scavenging activities of free radical by Nitraria tangutorun Bobr. red pigment in vitro. The article studied superoxide radical system, hydroxyl radical system and system of DPPH• and compared with Vc. It was showed that the scavenging effects appeared a ascending trend with the increasing of concentration of red pigment. And red pigment had different scavenging effects in three radical systems: 1)The scavenging capacity of red pigment in the hydroxyl radical and DPPH radical were relatively better than Vc (But their scavenging capacities were almost equal in higher concentration). 2)The scavenging capacity of red pigment in the superoxide radical system was lower than Vc and it was lower than 50% in selected concentration rage. In all, :Nitraria tangutorun Bobr. red pigment was a better kind of natural radical scavenger. 7. The animal experiments showed that anthocyanins from Nitraria tangutorum Bobr. fruit could effectively decrease blood lipids level of rats with hyperlipemia, improve blood lipids metabolism, reduce the formation of atherosclerosis, at the same time, effectively increase antioxidative level of rat body, improve antioxidase activities, reduce peroxidative lipid contents, and reduce oxidation response level. Main features were: they significantly decreased triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), apoprotein B(ApoB), atherosclerosis index(AI)TC-HDL-C/HDL-C and LDL-C/HDL-C (p<0.05), and significantly increased high density lipoprotein cholesterol (HDL-C) and apoprotein A (ApoA) level(p<0.05); in addition, they significantly increased TAC, superoxide dismutase(SOD) activities, decreased malonyldialdehyde(MDA) contents(p<0.05), and displayed significant dose-dependence. The blood lipids level and AIs of rats with hyperlipemia correlated closely with oxidation and antioxidation levels, both of which were cause and effect mutually and promoted each other. 8. By analyzing the quantity and effect relation and the structure and effect relation between anthocyanin compounds and blood lipids level and antioxidation, it brought forward the mechanism which black rice improves cardiovascular diseases by reducing serum lipid level and anti-atherosclerosis is the antioxidation and protection of its anthocyanin compounds for endothelial cells damaged by peroxidation. While the main mechanism for antioxidation of anthocyanin compounds of Nitraria tangutorum Bobr. fruit is that phenolic hydroxyl group provides hydrogen atom, prevents oxidation of body lipid and captures active free radicals and establishes phenol and quinone balance.
Pages111
Language中文
Document Type学位论文
Identifierhttp://210.75.249.4/handle/363003/3152
Collection中国科学院西北高原生物研究所
Recommended Citation
GB/T 7714
丁晨旭. 唐古特白刺(Nitraria tangutorum Bobr.)天然色素的研究[D]. 西北高原生物研究所. 中国科学院西北高原生物研究所,2008.
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