功能化磁性粒子—高效液相色谱分离分析体液中的单胺类神经递质
发布时间：2018-09-11 19:45
【摘要】：单胺类神经递质包括儿茶酚胺类的去甲肾上腺素(NE)、肾上腺素(E)和多巴胺(DA),以及吲哚胺类的5-羟色胺(5-HT)等,这些神经递质在生物体内起着重要作用,其含量与人体健康密切相关。由于体液组成复杂,且单胺类神经递质在体液中的含量低,因此,利用前处理过程对样品进行除杂与富集是非常必要的。与传统的样品前处理方法相比,功能化磁性粒子由于具有易于分离、安全无毒等优点,在医药、生物、环保等方面已得到广泛应用,但是将磁性粒子用于体液中单胺类神经递质分离分析方面的研究还很少。 本文成功地制备了三种不同功能化的磁性粒子,并利用多种手段对合成的功能化磁性粒子进行了表征,分析了它们的性质。分别利用这三种功能化磁性粒子对尿液中单胺类神经递质进行了分离分析研究,取得了令人满意的效果。 论文主要分为以下四个部分： 1.对单胺类神经递质、单胺类神经递质的检测方法、单胺类神经递质的前处理方法进行了综述,重点介绍了固相萃取在单胺类神经递质前处理方面的应用及功能化磁性粒子的制备。 2.采用多步法制备得到苯硼酸基功能化磁性纳米粒子(Fe3O4@APBA NPs),并通过透射电镜(TEM)、傅里叶红外光谱仪(FT-IR)、X射线衍射仪(XRD)和振动样品磁强计(VSM)等不同的表征手段对合成的Fe3O4@APBA NPs粒子进行表征。利用Fe3O4@APBA NPs对四种单胺类神经递质NE、E、DA及5-HT进行分离提取,结合高效液相色谱-电化学(HPLC-ECD)分析检测。在1mL料液中,确定了单胺类神经递质浓度为5μg/mL时,磁性粒子用量为10mg,萃取时间为5min,洗脱剂为0.02mol/L盐酸-甲醇的最佳萃取-洗脱条件。实验表明,Fe3O4@APBA NPs磁性粒子对三种儿茶酚胺类神经递质NE、E及DA均展示了较好的提取效果,而对吲哚胺类神经递质5-HT几乎没有提取,说明磁性粒子上的硼酸基对具有邻位酚羟基结构的化合物具有明显的选择性。在优化条件下,将该磁性粒子用于分离富集尿样中的NE、E和DA,结果显示,NE、E及DA在0.01-2.0μg/mL浓度范围内色谱峰高与浓度呈现良好的线性关系,检出限分别达到了7.8ng/mL、2.0ng/mL、7.9ng/mL,相对标准偏差均小于5.3%。将建立的方法用于人体尿样中单胺类神经递质的测定,获得了令人满意的结果,说明建立的Fe3O4@APBA NPs磁性粒子分离富集-高效液相色谱测定单胺类神经递质的方法是一种简单、快速、灵敏的新方法。 3.合成了二氧化钛包裹的介孔功能化磁性纳米粒子(Fe3O4@nSiO2@mTiO2NPs),并利用TEM、FT-IR、XRD、VSM和BET等表征手段对合成的粒子进行了表征分析。利用Fe3O4@nSiO2@mTiO2NPs对四种单胺类神经递质NE、E、DA及5-HT进行了分离提取,对料液相pH、提取时间、缓冲溶液种类与浓度、磁性粒子用量、洗脱溶剂种类与浓度等条件进行了优化。结果表明,合成的磁性粒子具有较高的选择性,对NE、E及DA展示了较好的萃取效果,而对5-HT几乎没有萃取。在优化条件下,该磁性粒子对NE、E和DA的富集因子范围在8-10之间,结合HPLC-ECD分析检测,在0.01-1.5μg/mL浓度范围内,NE、E和DA的峰高与浓度之间呈现良好的线性关系,检出限分别为11.8ng/mL、0.66ng/mL、6.0ng/mL。将建立的方法用于人体尿样中单胺类神经递质的测定,获得了令人满意的结果,说明建立的Fe3O4@nSiO2@mTiO2NPs磁性粒子分离富集-高效液相色谱测定单胺类神经递质的方法是一种简单、快速、灵敏的分析方法。 4.采用一步包埋法制备了羧甲基纤维素包裹的功能化磁性纳米粒子(Fe3O4@CMC NPs),并利用TEM、FT-IR、XRD、和VSM等手段对合成的磁性粒子进行了表征。利用Fe3O4@CMC NPs磁性粒子对四种单胺类神经递质NE、E、DA、5-HT进行分离提取,对料液相pH、提取时间、洗脱溶剂种类与浓度等条件进行了优化。在优化条件下,将该磁性粒子用于分离富集尿样中的单胺类神经递质,结合高效液相色谱-电化学方法(HPLC-ECD)分析检测,在线性范围为0.01-1.5μg/mL时,建立的方法对E和DA的检出限分别达到7.5和9.6ng/mL,说明建立的Fe3O4@CMC NPs磁性粒子分离富集-高效液相色谱法可以对尿样中的肾上腺素和多巴胺进行简单、快速、灵敏的分析。
[Abstract]:Monoamine neurotransmitters include catecholamine norepinephrine (NE), epinephrine (E), dopamine (DA), and indoleamine 5-hydroxytryptamine (5-HT). These neurotransmitters play an important role in organisms and their contents are closely related to human health. Because of the complex composition of humor and the presence of monoamine neurotransmitters in body fluid Compared with traditional sample pretreatment methods, functionalized magnetic particles have been widely used in medicine, biology, environmental protection and other fields because of their advantages of easy separation, safety and non-toxicity. However, magnetic particles are used in monoamine nerves in body fluids. There are few studies on the separation and analysis of transmitters.
In this paper, three kinds of magnetic particles with different functions were successfully prepared, and the synthesized magnetic particles were characterized by various means, and their properties were analyzed.
The thesis is mainly divided into four parts:
1. The methods for the detection of monoamine neurotransmitters, monoamine neurotransmitters and the pretreatment methods of monoamine neurotransmitters were reviewed. The application of solid phase extraction in the pretreatment of monoamine neurotransmitters and the preparation of functional magnetic particles were mainly introduced.
2. Phenylboronic acid-based functionalized magnetic nanoparticles (Fe3O4@APBA NPs) were prepared by multi-step method, and characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). Neurotransmitters NE, E, DA and 5-HT were separated and extracted. High performance liquid chromatography-electrochemical analysis (HPLC-ECD) was used to determine the optimum conditions for the Extraction-elution of monoamine neurotransmitters in 1 mL feed solution. When the concentration of monoamine neurotransmitters was 5 ug/mL, the dosage of magnetic particles was 10 mg, the extraction time was 5 min, and the eluent was 0.02 mol/L hydrochloric acid-methanol. APBA NPs magnetic particles showed good extraction effect on three catecholamine neurotransmitters NE, E and DA, but almost no extraction of indoleamine neurotransmitter 5-HT, indicating that the boric acid group on magnetic particles had obvious selectivity for compounds with o-phenolic hydroxyl structure. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentration in the range of 0.01-2.0 ug/mL. The detection limits were 7.8 ng/mL, 2.0 ng/mL and 7.9 ng/mL, respectively. The relative standard deviations were all less than 5.3%. The method was applied to the determination of monoamine neurotransmitters in human urine and the results were satisfactory. Satisfactory results showed that the method of separation and preconcentration of Fe3O4@APBA NPs magnetic particles and determination of monoamine neurotransmitters by high performance liquid chromatography was simple, rapid and sensitive.
3. Titanium dioxide-coated mesoporous functional magnetic nanoparticles (Fe3O4@nSiO2@mTiO2NPs) were synthesized and characterized by TEM, FT-IR, XRD, VSM and BET. Four monoamine neurotransmitters NE, E, DA and 5-HT were separated and extracted by Fe3O4@nSiO2@mTiO2NPs. The pH, extraction time, and extraction time were slowed down. The results showed that the magnetic particles had high selectivity and showed good extraction effect for NE, E and DA, but almost no extraction for 5-HT. Under the optimum conditions, the enrichment factor of the magnetic particles for NE, E and DA was in the range of 8-DA. The detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL respectively. The established method was applied to the determination of monoamine neurotransmitters in human urine with satisfactory results. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentrations in the range of 0.01-1.5 ug/mL, and the detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL, respectively. Separation and preconcentration of monoamine neurotransmitters with nSiO2@mTiO2NPs magnetic particles by high performance liquid chromatography is a simple, rapid and sensitive method.
4. Carboxymethylcellulose-coated functionalized magnetic nanoparticles (Fe3O4@CMC NPs) were prepared by one-step embedding method and characterized by TEM, FT-IR, XRD and VSM. Fe3O4@CMC NPs magnetic particles were used to separate and extract four monoamine neurotransmitters NE, E, DA, 5-HT, pH and extraction time. Under the optimized conditions, the magnetic particles were used to separate and enrich monoamine neurotransmitters in urine samples. The detection limits of E and DA were 7.5 and 9.6 n respectively in the linear range of 0.01-1.5 ug/ml by high performance liquid chromatography-electrochemical analysis (HPLC-ECD). G/mL, indicating that the established Fe3O4@CMC NPs magnetic particle separation and enrichment-high performance liquid chromatography can be used for the analysis of adrenaline and dopamine in urine samples simple, rapid and sensitive.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R917;O657.7
本文编号：2237687
[Abstract]:Monoamine neurotransmitters include catecholamine norepinephrine (NE), epinephrine (E), dopamine (DA), and indoleamine 5-hydroxytryptamine (5-HT). These neurotransmitters play an important role in organisms and their contents are closely related to human health. Because of the complex composition of humor and the presence of monoamine neurotransmitters in body fluid Compared with traditional sample pretreatment methods, functionalized magnetic particles have been widely used in medicine, biology, environmental protection and other fields because of their advantages of easy separation, safety and non-toxicity. However, magnetic particles are used in monoamine nerves in body fluids. There are few studies on the separation and analysis of transmitters.
In this paper, three kinds of magnetic particles with different functions were successfully prepared, and the synthesized magnetic particles were characterized by various means, and their properties were analyzed.
The thesis is mainly divided into four parts:
1. The methods for the detection of monoamine neurotransmitters, monoamine neurotransmitters and the pretreatment methods of monoamine neurotransmitters were reviewed. The application of solid phase extraction in the pretreatment of monoamine neurotransmitters and the preparation of functional magnetic particles were mainly introduced.
2. Phenylboronic acid-based functionalized magnetic nanoparticles (Fe3O4@APBA NPs) were prepared by multi-step method, and characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). Neurotransmitters NE, E, DA and 5-HT were separated and extracted. High performance liquid chromatography-electrochemical analysis (HPLC-ECD) was used to determine the optimum conditions for the Extraction-elution of monoamine neurotransmitters in 1 mL feed solution. When the concentration of monoamine neurotransmitters was 5 ug/mL, the dosage of magnetic particles was 10 mg, the extraction time was 5 min, and the eluent was 0.02 mol/L hydrochloric acid-methanol. APBA NPs magnetic particles showed good extraction effect on three catecholamine neurotransmitters NE, E and DA, but almost no extraction of indoleamine neurotransmitter 5-HT, indicating that the boric acid group on magnetic particles had obvious selectivity for compounds with o-phenolic hydroxyl structure. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentration in the range of 0.01-2.0 ug/mL. The detection limits were 7.8 ng/mL, 2.0 ng/mL and 7.9 ng/mL, respectively. The relative standard deviations were all less than 5.3%. The method was applied to the determination of monoamine neurotransmitters in human urine and the results were satisfactory. Satisfactory results showed that the method of separation and preconcentration of Fe3O4@APBA NPs magnetic particles and determination of monoamine neurotransmitters by high performance liquid chromatography was simple, rapid and sensitive.
3. Titanium dioxide-coated mesoporous functional magnetic nanoparticles (Fe3O4@nSiO2@mTiO2NPs) were synthesized and characterized by TEM, FT-IR, XRD, VSM and BET. Four monoamine neurotransmitters NE, E, DA and 5-HT were separated and extracted by Fe3O4@nSiO2@mTiO2NPs. The pH, extraction time, and extraction time were slowed down. The results showed that the magnetic particles had high selectivity and showed good extraction effect for NE, E and DA, but almost no extraction for 5-HT. Under the optimum conditions, the enrichment factor of the magnetic particles for NE, E and DA was in the range of 8-DA. The detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL respectively. The established method was applied to the determination of monoamine neurotransmitters in human urine with satisfactory results. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentrations in the range of 0.01-1.5 ug/mL, and the detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL, respectively. Separation and preconcentration of monoamine neurotransmitters with nSiO2@mTiO2NPs magnetic particles by high performance liquid chromatography is a simple, rapid and sensitive method.
4. Carboxymethylcellulose-coated functionalized magnetic nanoparticles (Fe3O4@CMC NPs) were prepared by one-step embedding method and characterized by TEM, FT-IR, XRD and VSM. Fe3O4@CMC NPs magnetic particles were used to separate and extract four monoamine neurotransmitters NE, E, DA, 5-HT, pH and extraction time. Under the optimized conditions, the magnetic particles were used to separate and enrich monoamine neurotransmitters in urine samples. The detection limits of E and DA were 7.5 and 9.6 n respectively in the linear range of 0.01-1.5 ug/ml by high performance liquid chromatography-electrochemical analysis (HPLC-ECD). G/mL, indicating that the established Fe3O4@CMC NPs magnetic particle separation and enrichment-high performance liquid chromatography can be used for the analysis of adrenaline and dopamine in urine samples simple, rapid and sensitive.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R917;O657.7
【参考文献】
相关期刊论文 前10条
1 邓向军;;高效液相-荧光检测法测定尿液中儿茶酚胺类化合物的含量[J];安徽医药;2007年01期
2 刘广印;沈佐君;何晓东;伊茂礼;;毛细管电泳-激光诱导荧光检测大鼠脑组织多巴胺含量[J];蚌埠医学院学报;2007年04期
3 李延年,伦立民,田青武,田润华,李晓菲,姜彩琴;尿中儿茶酚胺含量的高压液相色谱检测法[J];青岛大学医学院学报;2002年02期
4 洪小平,彭图治;功能高分子磁性微球的制备及分析应用[J];分析化学;2003年07期
5 马伟;孙登明;;聚L-精氨酸修饰电极存在下同时测定多巴胺和肾上腺素[J];分析化学;2007年01期
6 金谷;姚剑侠;姚奇志;;失水山梨醇脂肪酸酯-聚乙二醇嫁接的聚乙烯醇化磁性药物载体的制备及应用[J];分析化学;2010年05期
7 陈乙平;李顺兴;;纳米二氧化钛富集-分光光度法测定水体中痕量5-磺基水杨酸[J];分析化学;2010年11期
8 王贤亲;虞慧畅;林丹;张园;王学宝;;液相色谱-串联质谱法测定人血浆中儿茶酚胺的含量[J];分析试验室;2008年S1期
9 段语晖;卫引茂;;基于新型硼酸固相萃取柱的多巴胺色谱分析方法[J];分析化学;2013年03期
10 盖青青;屈锋;梅芳;张玉奎;;磁性粒子在蛋白质分离纯化中的应用[J];化学通报;2010年02期
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