重金属钯和汞的生物发光检测技术研究

发布时间：2018-03-06 10:20

  本文选题：重金属　切入点：生物发光　出处：《山东大学》2017年硕士论文　论文类型：学位论文

【摘要】：作为常见的污染物,重金属(Heavy Metal)是比重相对轻金属更大的一类金属,可以由工业污染、交通尾气以及食物、化妆品中引入。由于脂溶性高、易产生螯合作用等特性,重金属在自然界或人体内存在着蓄积的现象,当蓄积达到一定量时,容易引发机体各器官、各系统退行性功能损伤,因此危害巨大。除了传统的检测手段之外,最近新发展起来的荧光小分子探针检测具有较强的优势,但这些检测手段都存在无法应用于活体检测的缺点,因此本工作主要以重金属钯和汞为例,探索将前沿的生物发光检测手段应用于重金属的检测中。生物发光(Bioluminescence)是一种自然界普遍存在的,依赖于生物正常生命活动的化学发光。其本质是酶和底物之间的相互作用,经过一系列生物反应产生化学能,并以光能的形式释放出来。生物发光成像技术凭借其高灵敏性、高生物相容性以及可视化等特点已经成为很重要的检测手段,被广泛应用于各个领域。其中萤火虫生物发光系统是研究最为完善的一类生物发光系统。基于已报道的重金属钯和汞的荧光小分子探针的设计机理,本论文设计合成了两类五个生物发光探针,以开发用于重金属钯和汞的体内外检测技术。主要包括三个部分:一、前言;二、重金属钯的生物发光检测及其应用;三、重金属汞的生物发光检测及其应用。第一部分介绍了重金属的概况、其污染和毒性研究现状及传统检测方法;总结目前已有的重金属钯和汞的荧光小分子检测探针及其基本检测原理;概括了生物发光的原理及萤火虫萤光素系统。第二部分主要介绍了本课题研究的2个重金属钯生物发光探针。钯在珠宝装饰行业、交通行业、电子和电气行业、医疗行业以及工业催化行业均有相应的应用,但钯进入体内后会引起很大危害,因此本部分设计合成了两个生物发光钯探针。两个钯探针均以炔丙基醚基为水解基团,与钯作用后释放萤光素酶底物,从而实现检测钯的目的。该部分对探针的合成路线、体外应用、细胞水平应用及动物水平的应用进行了探讨。结果表明两个钯探针均可以实现体外对钯的线性检测,但CH-1的强度较弱,因此在动物水平仅以CH-2为检测探针,结果表明该探针可以实现对钯在小鼠体内的检测。第三部分主要介绍了本课题研究的3个汞生物发光探针。汞是一种非常重要的重金属,在各个行业领域中都有应用,包括制造行业、医药行业、化妆品行业以及矿产业等。汞是一中毒性非常大的重金属,临床上重金属中毒的案例不在少数,而且随着工业发展,汞的大规模污染也有发生,造成很大的危害,因此本部分设计合成了三个汞离子探针。分别以乙烯基醚基和丁炔基醚基为水解基团,在汞的作用下释放萤光素酶底物,用以检测汞。该部分对三个探针的合成路线,体外检测,细胞水平应用以及动物水平的应用进行了探讨。结果表明,CHG-1由于反应温度原因,只能实现体外检测,而CHG-3检测范围窄,灵敏性差,因此选择检测范围窄且易于反应的CHG-2作为体内水平检测的探针,CHG-2体内可以实现对汞中毒的检测,而且有对汞中毒体内蓄积状态进行检测的潜在价值。
[Abstract]:As the common pollutants, heavy metals (Heavy, Metal) is a kind of light metal metal proportion is relatively larger, from industrial pollution, traffic exhaust and food, cosmetics was introduced. Due to the high fat soluble, easy to produce the effect of chelating characteristics of heavy metals in the accumulation phenomenon in nature or human memory, when the accumulation reached a certain amount of time, easy to cause the body organs function, degenerative system damage, so the damage is huge. In addition to the traditional means of detection, fluorescent molecular probe detection recently developed has a strong advantage, but these measures cannot be applied to in vivo detection there are shortcomings, so this work is mainly to heavy metal palladium and mercury as an example, to explore the frontiers of bioluminescence detection methods in the detection of heavy metal application. Bioluminescence (Bioluminescence) is a kind of universal existence in nature, depends on the biological Chemical normal life activity light. Its essence is the interaction between the enzyme and the substrate, after a series of biological reaction and chemical energy, light energy is released. Bioluminescence imaging technology with its high sensitivity, high biocompatibility and visualization features detection means has become very important and widely used in various fields. The firefly bioluminescence system is a kind of biological sound emitting system as the research design. The mechanism of the fluorescence molecular probes of heavy metals have been reported palladium and mercury based on two kinds of five bioluminescent probes were designed and synthesized in this paper, in order to develop for heavy metal palladium and mercury in vivo detection technology mainly includes three parts: first, introduction; two, heavy metal palladium bioluminescence detection and its application; three, mercury bioluminescence detection and its application. The first part is the introduction of a large amount of money The general situation, the pollution and toxicity research and traditional detection methods; summarizes the current detection of fluorescent molecules for heavy metal palladium and mercury probe and its basic detection principle; summarizes the principle and firefly bioluminescence in fluorescent system. The second part mainly introduces the research of 2 metal palladium bioluminescence probe. Palladium traffic in the jewelry industry, decoration industry, electronic and electrical industry, medical industry and industrial application of catalytic industry has a corresponding, but palladium enters the body will cause great harm, so the design and synthesis of two bioluminescent probes. Two palladium palladium probe with propargyl ether base for the hydrolysis of groups with palladium prime Enzyme Fluorescence liberating effect, so as to achieve the purpose of detection PD. The part of synthetic routes of the probe in vitro application, application level and application level of animal cells is discussed. The results show that the two palladium probe can achieve linear for PD detection in vitro, but weaker CH-1, so in the animal level only using CH-2 as the detection probe, the results show that the probe can be achieved on the palladium in the detection of mice. The third part mainly introduces the research of 3 mercury biological luminescent probe. Mercury is a kind of heavy metal is very important, are widely used in various industries, including manufacturing industry, pharmaceutical industry, cosmetics industry and mining industry. Mercury is very toxic heavy metals in clinical heavy metal poisoning in a few cases, but with the development of industry, large-scale pollution of Mercury also can occur. Great harm, so the synthesis of three mercury ion probe design. This part respectively with vinyl ether and ether groups for the hydrolysis of Ding Queji, in the role of mercury release luciferase substrate for detection of mercury. 璇ラ儴鍒嗗�逛笁涓�鎺㈤拡鐨勫悎鎴愯矾绾，

本文编号：1574432