基于主客体反应的微囊藻毒素纯化与检测方法的研究
发布时间：2019-02-18 15:05
【摘要】：微囊藻毒素能够引发原发性肝癌和大肠癌,其在饮用水和蔬菜鱼和贝类等食品中的出现,引发了人类对微囊藻毒素(MC-LR)越来越多的关注,因此需要一种快速高效简便精确和低廉的检测微囊藻毒素的方法。但是样品中微囊藻毒素的浓度一般较低,并且存在许多其它复杂的成分干扰测定,在测定微囊藻毒素之前,需要对样品进行预处理,得到浓度较高的微囊藻毒素样品。本研究首先用单-(6-巯基)β-环糊精和巯基磷酸双修饰纳米金,再用红外光谱全波长扫描和琼脂糖凝胶电泳分别表征单-(6-巯基)β-环糊精和和修饰的纳米金。结果表明对纳米金成功地进行了双修饰,双修饰的纳米金能够在体系中稳定存在并且对微囊藻毒素具有良好的吸附性能,其吸附量高达2.125μg/mL,静态分布系数为5.666。因此通过藻毒素结构中酪氨酸残基与β-环糊精,及其精氨酸残基与磷酸的主客体相互作用可以实现藻毒素的特异性识别。基于纳米金双修饰后对微囊藻毒素表现出的良好吸附性,本研究以壳聚糖为载体进行了磷酸和环糊精双修饰(CTS-CD-P),制备了一种高效高特异的固相萃取材料,通过红外光谱表征了合成材料,验证了化学修饰的成功。通过高效液相色谱(HPLC)测定了其对微囊藻毒素(MC-LR)的吸附量。结果表明,双修饰的壳聚糖(CTS-CD-P)对MC-LR有较高的吸附率,溶解和固态状态CTS-CD-P对MC-LR吸附量最高分别为9.04μmol/g和3.40μmol/g。本研究基于双修饰纳米金的良好的吸附性能建立了一种基于微分脉冲伏安分析的电化学微藻毒素检测方法。首先通过循环伏安法制备中性红聚合修饰的玻碳电极,利用中性红聚合物表面的正电荷吸附巯基-β-环糊精和巯基磷酸修饰双修饰纳米金,修饰好的电极于磷酸缓冲液中测定藻毒素浓度,结果表明修饰电极对MC-LR的线性范围为:26nM-213μM,检测限为(3σ/S)19nM,其与高效液相色谱法的检测限接近。
[Abstract]:Microcystins can cause primary liver cancer and colorectal cancer, its in drinking water and vegetables? The emergence of food such as fish and shellfish has attracted more and more attention to microcystins (MC-LR) and therefore requires a rapid response. Efficient? Easy? An accurate and inexpensive method for the detection of microcystins. However, the concentration of microcystins in the samples is generally low, and there are many other complex components interference determination. Before the determination of microcystins, the samples need to be pretreated to get the higher concentrations of microcystins. In this study, nanocrystalline gold was modified with mono- (6-mercaptoyl) 尾 -cyclodextrin and mercaptophosphoric acid. The single (6-mercapto) 尾 -cyclodextrin and modified gold nanoparticles were characterized by full-wavelength scanning and agarose gel electrophoresis, respectively. The results showed that the nanocrystalline gold was successfully modified. The nanocrystalline gold could exist stably in the system and had good adsorption performance for microcystins. The adsorption capacity was up to 2.125 渭 g / mL, and the static distribution coefficient was 5.666. Therefore, the specific recognition of phycotoxin can be achieved by the interaction of tyrosine residue with 尾 -cyclodextrin and the host and guest interaction of arginine residue and phosphoric acid in the structure of phycotoxin. Based on the good adsorption of microcystins by gold nanoparticles, a high efficient and specific solid phase extraction material was prepared by phosphoric acid and cyclodextrin modification (CTS-CD-P) with chitosan as the carrier. The synthesized materials were characterized by IR spectra, and the success of the chemical modification was verified. The adsorption capacity of microcystin (MC-LR) was determined by high performance liquid chromatography (HPLC). The results show that the double modified chitosan (CTS-CD-P) has a high adsorption rate for MC-LR, and the highest adsorption capacity for MC-LR by dissolved and solid CTS-CD-P is 9.04 渭 mol/g and 3.40 渭 mol/g., respectively. Based on the good adsorption properties of double modified gold nanoparticles, a new method for the detection of microalgae toxins based on differential pulse voltammetry (DPA) was developed. Firstly, the glassy carbon electrode modified by neutral red polymerization was prepared by cyclic voltammetry. The adsorption of thiol-尾-cyclodextrin and sulfhydryl phosphoric acid on the surface of neutral red polymer was used to modify gold nanoparticles. The modified electrode was used to determine the concentration of phycotoxin in phosphoric acid buffer. The results showed that the linear range of modified electrode for MC-LR was 26nM-213 渭 M, the detection limit was (3 蟽 / S) 19 nm, which was close to the detection limit of high performance liquid chromatography (HPLC).
【学位授予单位】：中南民族大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O657.1
，
本文编号：2425958
[Abstract]:Microcystins can cause primary liver cancer and colorectal cancer, its in drinking water and vegetables? The emergence of food such as fish and shellfish has attracted more and more attention to microcystins (MC-LR) and therefore requires a rapid response. Efficient? Easy? An accurate and inexpensive method for the detection of microcystins. However, the concentration of microcystins in the samples is generally low, and there are many other complex components interference determination. Before the determination of microcystins, the samples need to be pretreated to get the higher concentrations of microcystins. In this study, nanocrystalline gold was modified with mono- (6-mercaptoyl) 尾 -cyclodextrin and mercaptophosphoric acid. The single (6-mercapto) 尾 -cyclodextrin and modified gold nanoparticles were characterized by full-wavelength scanning and agarose gel electrophoresis, respectively. The results showed that the nanocrystalline gold was successfully modified. The nanocrystalline gold could exist stably in the system and had good adsorption performance for microcystins. The adsorption capacity was up to 2.125 渭 g / mL, and the static distribution coefficient was 5.666. Therefore, the specific recognition of phycotoxin can be achieved by the interaction of tyrosine residue with 尾 -cyclodextrin and the host and guest interaction of arginine residue and phosphoric acid in the structure of phycotoxin. Based on the good adsorption of microcystins by gold nanoparticles, a high efficient and specific solid phase extraction material was prepared by phosphoric acid and cyclodextrin modification (CTS-CD-P) with chitosan as the carrier. The synthesized materials were characterized by IR spectra, and the success of the chemical modification was verified. The adsorption capacity of microcystin (MC-LR) was determined by high performance liquid chromatography (HPLC). The results show that the double modified chitosan (CTS-CD-P) has a high adsorption rate for MC-LR, and the highest adsorption capacity for MC-LR by dissolved and solid CTS-CD-P is 9.04 渭 mol/g and 3.40 渭 mol/g., respectively. Based on the good adsorption properties of double modified gold nanoparticles, a new method for the detection of microalgae toxins based on differential pulse voltammetry (DPA) was developed. Firstly, the glassy carbon electrode modified by neutral red polymerization was prepared by cyclic voltammetry. The adsorption of thiol-尾-cyclodextrin and sulfhydryl phosphoric acid on the surface of neutral red polymer was used to modify gold nanoparticles. The modified electrode was used to determine the concentration of phycotoxin in phosphoric acid buffer. The results showed that the linear range of modified electrode for MC-LR was 26nM-213 渭 M, the detection limit was (3 蟽 / S) 19 nm, which was close to the detection limit of high performance liquid chromatography (HPLC).
【学位授予单位】：中南民族大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O657.1
，
本文编号：2425958
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