多酸纳米粒子在药物及蛋白质检测中的应用研究
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摘要
多酸化合物作为一种独特的金属氧化物具有很多特殊性质。基于这些性质,多酸化合物在催化剂,抗肿瘤药物以及作为分析检测探针等生化研究领都拥有优良的表现,也因此吸引了众多研究者的关注。随着纳米技术的发展,多酸化合物的合成和应用也进入纳米领域。目前,已经被报道的多酸化合物的纳米微粒涵盖了多种形态,包括纳米粒子,纳米棒,纳米线,纳米管,纳米多孔材料等;而合成方法也是多种多样的,已有的方法包括模版合成,气-液-固多相合成,电化学加工,液相合成等。其中,液相合成是研究较多的一种方法。这些多酸的纳米材料被用于催化剂及药物研究,都表现出了更优异的特质。已有的报道表明,多酸化合物除了可以用来检测微量痕量金属离子之外,还可以直接用作分析试剂来检测药物及蛋白质等大分子。而这些研究都是直接使用了多酸化合物分子,利用多酸的纳米粒子作为分析探针的研究极少。本文将两种不同的多酸纳米材料作为分析检测探针,应用到药物和蛋白质的检测中去。首先,以四氯化碘钼酸盐与结晶紫的复合纳米颗粒为探针,发展了一种用紫外可见吸收光度法检测多巴胺的快捷灵敏的方法;其次,探讨了一种Keggin型多酸纳米粒子Ag_3PW_(l2)O_4的液相合成方法,并以其为探针用光散射的方法测定了朊蛋白、溶菌酶以及多巴胺。具体研究内容包括以下两个大方面:1.以四氯化碘钼酸盐与结晶紫的复合纳米颗粒为探针,发展了一种用紫外可见吸收光度法检测多巴胺的快捷灵敏的方法。在酸性条件下,随着多巴胺浓度的增加,富含纳米粒子溶液的颜色也随之加深,可以建立一种可视化半定量检测方法。在紫外可见分光光度计下得到多巴胺的线性关系在5.0x10~(-7)-1.4×10~(-5) mol·L-~浓度范围内,检出限(3σ)为9.0x10~(-8) mol·L~(-1)。该方法被成功应用于盐酸多巴胺注射液的测定。2.研究了一种Ag_3PW_(l2)O_4纳米立方体的可控合成路线。并且近一步以该纳米粒子为分析探针,利用光散射信号检测了蛋白质及药物小分子。(1).研究发现,在有机小分子二甲基甲酰胺(DMF)的调控作用下Keggin型多酸化合物Ag_3PW_(l2)O_4在水溶液中形成规则的纳米立方体,随温度改变该纳米立方体转变为空心立方结构。且基于该多酸纳米粒子,我们建立了一种以光散射信号为表征手段的检测朊蛋白的新方法。该多酸纳米立方体可以与朊蛋白(PrP~C)相互作用,产生显著的光散射信号增强,其增强程度与36-840 ng·mL~(-1)浓度范围的朊蛋白呈线性关系。并且在相同实验条件下,与其它多种蛋白相比,与朊蛋白的相互作用表现出较好的选择性。(2).实验发现,多酸纳米粒子Ag_3PW_(l2)O_(40)可以与溶菌酶相互作用引起光散射信号的增强,建立了一种溶菌酶的光散射测定方法。在磷酸缓冲溶液pH 6.8的条件下,光散射信号在溶菌酶浓度为0.5-140μg·mL~(-1)。的范围内呈线性增强,检出限(3σ)为0.04μg·mL~(-1)。这个方法成功用于合成样中溶菌酶的测定。(3).研究表明,在醋酸一醋酸钠缓冲溶液pH 5.0的条件下,多酸纳米粒子Ag_3PW_(l2)O_(40)可以与多巴胺相互作用。以光散射信号为表征手段,可以得出在多巴胺浓度为3.0xl0~(-8)-1.2x10~(-5)mol·L~(-1)的范围内光散射信号随着多巴胺浓度的增加呈线性增强,检出限(3σ)为1.4x10~(-8)mol·L~(-1)。这种多巴胺的光散射测定方法被成功应用于盐酸多巴胺注射液的测定。
Polyoxometalates (POM) have a lot of unique properties as a kind of special metallic oxides. Because of these properties, they played an important role in a variety of field, such as catalysis, anticancer drugs and analytical reagents, all of these have attracted much attention by researchers. With the development of nanotechnology, the synthesis and application of POM has come into nanometer field. At present, many kinds of POM nanoparticles have already been reported, including nanoball, nanorod, nanowire, nanotube and the nanohollow on the film. And all these POM nanoparticles prepared by using a lot of different ways, such as template-assisted, electrochemical processes, vapor-liquid-solids-assisted and liquid-assisted. As a sort of analytical reagent, the POM could be used to determine the metal ion, some medicament and protein molecule. But most of these have been determined by using POM molecule not the POM nanoparticles. In this paper, two kinds of POM nanoparticles have been prepared and applied to analyse medicament and protein by UV and light scattering signals. The main content is as follows:
1. A spectrophotometric method for the determination of dopamine is described based on a type of ployoxometalates-dye complex nanoparticles which are composed by crystal violet with molybdate-iodine tetrachloride. By the reaction between dopamine and molybdate-iodine tetrachloride reagent in acidic medium, the color of the solution has changed, and an analysis method of dopamine could be constructed. By spectrophotometric signal, a linear relationship has been obtained when the concentration of dopamine is in the range of 5.0×10~(-7)~1.4×l0~(-5) mol·L~(-1), and the detection limit is 9.0×l0~(-8) mol·L~(-1) (3σ). This method has been used for analysis of the injection containing dopamine hydrochloride.
2. A controllable synthetic route of the Ag_3PW_(12)O_(40) nanocubes was investigated on our work, and these prepared nanocubes were used as an analysis probe to detect the protein and medicament.(1). It was found in this contribution that Keggin-type of polyoxometalates nanocubes could be formed in aqueous medium under the adjustment of dimethylformamide (DMF), which could be changed from solid nanocubes to hollow ones with the increasing of temperature. Further investigations showed that this Keggin-type of polyoxometalates nanocubes can specifically interact with prion protein, and the light scattering signals were enhanced in proportional to the content of prion in the range of 3.36-840 ng·mL~(-1).(2). The interaction of Ag_3PW_(12)O_(40) nanocubes and lysozyme was studied with light scattering (LS) spectral. Ag_3PW_(12)O_(40) nanocubes can be linked with lysozyme, and strongly enhanced their LS intensity. In the PB buffer solution with pH value of 6.8, the enhancing LS intensity was in proportion to the concentration of lysozyme in a range of 0.5-140μg·mL~(-1). The effects of reaction time, pH of solution, coexisted ions and organic regents were studied in this work. Under optimum conditions, the detection limit of lysozyme (3σ) is 0.04μg·mL~(-1) with nanoparticles as a probe. This method was successfully used to the detection of trace lysozyme.(3). A light scattering (LS) method for the determination of dopamine is described based on Ag_3PW_(12)O_(40) nanocubes. The dopamine can interact with Ag_3PW_(12)O_(40) nanocubes, and their LS signal was strongly enhanced. In the CH_3COOH-CH_3COONa buffer solution with pH value of 5.0, we can determine dopamine in a range of 3.0×10~(-8)-1.2×l0~(-5) mol·L~(-1), and the limit of determination (3σ) is 1.4×10~(-8) mol·L~(-1). The recommended method has been used for analysis of the injection containing dopamine hydrochloride.
Polyoxometalates (POM) have a lot of unique properties as a kind of special metallic oxides. Because of these properties, they played an important role in a variety of field, such as catalysis, anticancer drugs and analytical reagents, all of these have attracted much attention by researchers. With the development of nanotechnology, the synthesis and application of POM has come into nanometer field. At present, many kinds of POM nanoparticles have already been reported, including nanoball, nanorod, nanowire, nanotube and the nanohollow on the film. And all these POM nanoparticles prepared by using a lot of different ways, such as template-assisted, electrochemical processes, vapor-liquid-solids-assisted and liquid-assisted. As a sort of analytical reagent, the POM could be used to determine the metal ion, some medicament and protein molecule. But most of these have been determined by using POM molecule not the POM nanoparticles. In this paper, two kinds of POM nanoparticles have been prepared and applied to analyse medicament and protein by UV and light scattering signals. The main content is as follows:
1. A spectrophotometric method for the determination of dopamine is described based on a type of ployoxometalates-dye complex nanoparticles which are composed by crystal violet with molybdate-iodine tetrachloride. By the reaction between dopamine and molybdate-iodine tetrachloride reagent in acidic medium, the color of the solution has changed, and an analysis method of dopamine could be constructed. By spectrophotometric signal, a linear relationship has been obtained when the concentration of dopamine is in the range of 5.0×10~(-7)~1.4×l0~(-5) mol·L~(-1), and the detection limit is 9.0×l0~(-8) mol·L~(-1) (3σ). This method has been used for analysis of the injection containing dopamine hydrochloride.
2. A controllable synthetic route of the Ag_3PW_(12)O_(40) nanocubes was investigated on our work, and these prepared nanocubes were used as an analysis probe to detect the protein and medicament.(1). It was found in this contribution that Keggin-type of polyoxometalates nanocubes could be formed in aqueous medium under the adjustment of dimethylformamide (DMF), which could be changed from solid nanocubes to hollow ones with the increasing of temperature. Further investigations showed that this Keggin-type of polyoxometalates nanocubes can specifically interact with prion protein, and the light scattering signals were enhanced in proportional to the content of prion in the range of 3.36-840 ng·mL~(-1).(2). The interaction of Ag_3PW_(12)O_(40) nanocubes and lysozyme was studied with light scattering (LS) spectral. Ag_3PW_(12)O_(40) nanocubes can be linked with lysozyme, and strongly enhanced their LS intensity. In the PB buffer solution with pH value of 6.8, the enhancing LS intensity was in proportion to the concentration of lysozyme in a range of 0.5-140μg·mL~(-1). The effects of reaction time, pH of solution, coexisted ions and organic regents were studied in this work. Under optimum conditions, the detection limit of lysozyme (3σ) is 0.04μg·mL~(-1) with nanoparticles as a probe. This method was successfully used to the detection of trace lysozyme.(3). A light scattering (LS) method for the determination of dopamine is described based on Ag_3PW_(12)O_(40) nanocubes. The dopamine can interact with Ag_3PW_(12)O_(40) nanocubes, and their LS signal was strongly enhanced. In the CH_3COOH-CH_3COONa buffer solution with pH value of 5.0, we can determine dopamine in a range of 3.0×10~(-8)-1.2×l0~(-5) mol·L~(-1), and the limit of determination (3σ) is 1.4×10~(-8) mol·L~(-1). The recommended method has been used for analysis of the injection containing dopamine hydrochloride.
引文
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