量子点做探针共振瑞利散射法在多糖分析中的应用研究
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摘要
量子点,又称半导体纳米晶,三维尺度在1-10nm。由于他们特殊的发光性能,已经引起了广泛关注。量子点作为探针,已经被用于生物大分子,金属离子,药物等的分析测定。因此,量子点在新的分析测定领域具有很好的发展前景。
共振瑞利散射(resonance Rayleigh scattering, RRS)技术是二十世纪九十年代发展起来的一门新技术,由于其高的灵敏度、较好的选择性和简易性而引起科研工作者的兴趣和关注,近年来,已在核酸、蛋白质等生物大分子、药物、以及金属和非金属离子等的测定中得到很多的应用。最新研究表明,它除了上述应用外,对于研究纳米微粒也是一种有用的技术。而且利用量子点做探针共振瑞利散射法测定多糖的报道较少。
本论文分为两部分,第一部分是文献综述,主要介绍了量子点的制备和应用,共振瑞利散射技术及其发展以及多糖的介绍和常用分析方法。第二部分是研究报告,利用量子点做探针用共振瑞利散射光谱法分析和测定两种蛋白多糖。
一、硫酸皮肤素的测定
1、硒化镉量子点与硫酸皮肤素作用的共振瑞利散射光谱法研究
本文以巯基乙胺(2-mercaptoethylamine hydrochloride, CA)为稳定剂在水相中合成了2nm左右的CdSe量子点(CdSe quantum dots, CdSe QDs),基于硫酸皮肤素(Dermatan Sulfate, DS)所引起的CdSe QDs共振瑞利散射的增强,本文提出了一种检测硫酸皮肤素(Dermatan Sulfate, DS)的新方法。在最优反应条件下,RRS增强的强度与DS的浓度在一定范围内成正比。RRS的检出限为1.3ng·mL-1。讨论了实验最佳条件和共存物质对体系的影响,结果显示该方法具有较好的选择性。该方法用于人血清与尿样中DS的检测,结果另人满意。此外,结合紫外吸收光谱和透射电子显微镜(TEM)技术对共振瑞利散射光谱的增强的机理进行了探讨。据此建立了利用CdSe量子点测定DS的新方法。
2、CdTe量子点-硫酸皮肤素体系的共振瑞利散射光谱及其分析应用
本文以巯基乙胺为稳定剂在水相中合成了粒径很小的CdTe量子点(QDs)。采用共振瑞利散射光谱法,研究了CA-CdTe QDs与硫酸皮肤素(DS)的相互作用。CA-CdTe QDs与DS的相互作用导致了CA-CdTe QDs-DS体系RRS的显著增强,散射强度(ΔIRRS)与DS的浓度在一定范围内成正比,其线性范围和检出限分别为0.0039～0.6μg·mL-1和1.2 ng·mL-1,据此建立了CA-CdTe QDs作探针RRS法检测DS的高灵敏度新方法。同时讨论了最佳的反应条件和影响因素,研究了该方法的选择性。此外还讨论了RRS增强的原因。实验表明该方法有较高的灵敏度,应用于血样和尿样中样品的测定,结果令人满意。
二、葡聚糖硫酸钠的测定
1、硒化镉量子点做探针共振瑞利光谱法测定葡聚糖硫酸钠
以巯基乙胺为稳定剂在水相中合成CdSe量子点,在乙酸-乙酸钠缓冲溶液中,巯基乙胺自组装在量子点的表面形成带有正电的聚集体可与带负电的葡聚糖硫酸钠(Dextran Sulfate Sodium, DSS)靠静电引力及疏水作用力结合形成粒径较大的聚集体,这种聚集体的形成导致共振瑞利散射强度显著增大,最大散射峰位于536nm。建立了用CdSe量子点作为探针RRS法测定葡聚糖硫酸钠的灵敏度高,简便,快速的新方法,同时讨论了最佳反应条件和影响因素,该方法具有较好的选择性。应用于血样和尿样的测定结果令人满意。
2、CdTe量子点-葡聚糖硫酸钠体系共振瑞利散射光谱研究及其分析应用
合成了巯基乙氨为稳定剂的CdTe量子点(QDs)。利用吸收光谱和共振瑞利散射光谱(RRS)研究了CdTe QDs与葡聚糖硫酸钠(DSS)的相互作用。CdTe QDs与DSS的吸收光谱在反应前后发生了明显的变化,这说明CdTe QDs与DSS之间存在着强烈的相互作用。在pH 5.0的缓冲溶液中,CdTe QDs-DSS体系的结合前后RRS变化显著。据此推测DSS通过静电引力和疏水作用力吸附到CdTe QDs表面形成了结合产物,导致了CdTe QDs-DSS体系的RRS显著增强。研究了最佳的反应条件和影响因素,同时研究了该方法的选择性。应用于血样和尿样的测定,结果令人满意。
Quantum dots (QDs), which are semiconductor nanoparticles that have all three dimensions confined to the 1-10 nm length scale, have attracted considerable attention because of their unique luminescent properties. QDs have been applied to the detection of biomacromolecules, metal ions, pharmaceuticals, and so on. Therefore, it is significant for further developing new analytical application of QDs.
Resonance Rayleigh scattering (RRS) technology was developed during 1990s. As a newly analytical technique, it has been received much attention because of its sensitivity and simplicity Therefore, this technique has been increasingly applied to the determination of macromolecules such as inorganic ions, organic compounds, nucleic acids, proteins, cationic surfactant, and some drugs. In recent years, this technique was applied to study nanometer particle. However, QDs were used as probes for the determination of polysaccharides by resonance Rayleigh scattering has few reports.
This paper contained two parts. The first part was the literature review and it includse the preparation and application of quantum dots, the development of resonance Rayleigh scattering technology and the introduction of polysaccharides in common analysis. The second part was research paper. In this paper, quantum dots were used as probes for the determination of two proteoglycans by resonance Rayleigh scattering method.
Determination of Dermatan Sulfate:
1. Resonance Rayleigh scattering method for the determination of Dermatan
Sulfate with CdSe quantum dots as probes
The CdSe quantum dots capped by 2-mercaptoethylamine hydrochloride with an average diameter of 2 rm were synthesized in water solution. It could react with dermatan sulfate (DS) and form the large aggregates by virtue of electrostatic attraction and the hydrophobic force, which induced a great enhancement of RRS. In a certain ranges, the enhancements of scattering intensity (ΔI) were directly proportional to the concentration of DS. This method revealed good sensitiviy, and the detection limit (38) was 1.3 ng mL-1. The optimum conditions of this reaction system and the influencing factors were investigated. In addition, ultraviolet absorption spectra and transmission electron microscope were employed to discuss the reaction mechanism of RRS enhancement. A novel method for the determination of trace DS using CdSe quantum dots probes was developed.
2. Resonance Rayleigh scattering spectra of CdTe QDs-DS system and its analytical application
The CdTe quantum dots capped by 2-mercaptoethylamine hydrochloride was synthesized in water solution. CdTe QDs can react with dermatan sulfate and form an ion association complex. The linear relactionship between the relative intensity ofΔIRRS and concentration of DS was in the range of 0.2-0.6μg mL-1. A sensitive, simple, and selective method for the determination of DS with CdTe QDs as probe by resonance Rayleigh scattering technique was developed. The optimum conditions and the influence factors were investigated. Moreover, the composition of ion-association complexes and the reaction mechanism were discussed. The method was applied to the determination of DS in human blood with satisfactory results.
Determination of Dextran Sulfate Sodium:
1. Resonance Rayleigh scattering method for the determination of dermatan sulfate sodium with water solubility CdSe quantum dots as probe
In acetic acid-sodium acetate buffer medium, the surface of QDs was assembled by 2-mercaptoethylamine hydrochloride which showed positive charge. The CdSe QDs could react with dextran sulfate sodium (DSS) and form the large aggregates by virtue of electrostatic attraction and the hydrophobic force, which leads to the remarkable enhancement of RRS. The maximum scattering peak was located at 343 nm. A sensitive and simple method for the determination of DSS with CdSe QDs as probes by resonance Rayleigh scattering technique was developed. The optimum conditions and influence factors of the reactions were investigated. It has been applied to the determination of DSS in human blood and urine samples and this method exhibited satisfactory results.
2. Resonance Rayleigh scattering spectra of CdTe QDs-DSS system and its analytical application
2-Mercaptoethylamine hydrochloride capped CdTe quantum dots with the diameter of 2-3 nm were synthesized in aqueous solution. At pH 5.0, CdTe QDs could react with DSS and form the large aggregates by electrostatic attraction and the hydrophobic force, which resulted in a great enhancement of resonance Rayleigh scattering. In this paper, the relationship between ultraviolet-visible absorption and resonance Rayleigh scattering was studied. The affecting factors and characteristics of RRS for the interaction of CdTe QDs with DSS were also investigated and a sensitive, simple and fast method for the determination of trace amounts of DSS using the RRS method was developed. It has been applied to the determination of DSS in human blood and urine samples with satisfactory results.
共振瑞利散射(resonance Rayleigh scattering, RRS)技术是二十世纪九十年代发展起来的一门新技术,由于其高的灵敏度、较好的选择性和简易性而引起科研工作者的兴趣和关注,近年来,已在核酸、蛋白质等生物大分子、药物、以及金属和非金属离子等的测定中得到很多的应用。最新研究表明,它除了上述应用外,对于研究纳米微粒也是一种有用的技术。而且利用量子点做探针共振瑞利散射法测定多糖的报道较少。
本论文分为两部分,第一部分是文献综述,主要介绍了量子点的制备和应用,共振瑞利散射技术及其发展以及多糖的介绍和常用分析方法。第二部分是研究报告,利用量子点做探针用共振瑞利散射光谱法分析和测定两种蛋白多糖。
一、硫酸皮肤素的测定
1、硒化镉量子点与硫酸皮肤素作用的共振瑞利散射光谱法研究
本文以巯基乙胺(2-mercaptoethylamine hydrochloride, CA)为稳定剂在水相中合成了2nm左右的CdSe量子点(CdSe quantum dots, CdSe QDs),基于硫酸皮肤素(Dermatan Sulfate, DS)所引起的CdSe QDs共振瑞利散射的增强,本文提出了一种检测硫酸皮肤素(Dermatan Sulfate, DS)的新方法。在最优反应条件下,RRS增强的强度与DS的浓度在一定范围内成正比。RRS的检出限为1.3ng·mL-1。讨论了实验最佳条件和共存物质对体系的影响,结果显示该方法具有较好的选择性。该方法用于人血清与尿样中DS的检测,结果另人满意。此外,结合紫外吸收光谱和透射电子显微镜(TEM)技术对共振瑞利散射光谱的增强的机理进行了探讨。据此建立了利用CdSe量子点测定DS的新方法。
2、CdTe量子点-硫酸皮肤素体系的共振瑞利散射光谱及其分析应用
本文以巯基乙胺为稳定剂在水相中合成了粒径很小的CdTe量子点(QDs)。采用共振瑞利散射光谱法,研究了CA-CdTe QDs与硫酸皮肤素(DS)的相互作用。CA-CdTe QDs与DS的相互作用导致了CA-CdTe QDs-DS体系RRS的显著增强,散射强度(ΔIRRS)与DS的浓度在一定范围内成正比,其线性范围和检出限分别为0.0039～0.6μg·mL-1和1.2 ng·mL-1,据此建立了CA-CdTe QDs作探针RRS法检测DS的高灵敏度新方法。同时讨论了最佳的反应条件和影响因素,研究了该方法的选择性。此外还讨论了RRS增强的原因。实验表明该方法有较高的灵敏度,应用于血样和尿样中样品的测定,结果令人满意。
二、葡聚糖硫酸钠的测定
1、硒化镉量子点做探针共振瑞利光谱法测定葡聚糖硫酸钠
以巯基乙胺为稳定剂在水相中合成CdSe量子点,在乙酸-乙酸钠缓冲溶液中,巯基乙胺自组装在量子点的表面形成带有正电的聚集体可与带负电的葡聚糖硫酸钠(Dextran Sulfate Sodium, DSS)靠静电引力及疏水作用力结合形成粒径较大的聚集体,这种聚集体的形成导致共振瑞利散射强度显著增大,最大散射峰位于536nm。建立了用CdSe量子点作为探针RRS法测定葡聚糖硫酸钠的灵敏度高,简便,快速的新方法,同时讨论了最佳反应条件和影响因素,该方法具有较好的选择性。应用于血样和尿样的测定结果令人满意。
2、CdTe量子点-葡聚糖硫酸钠体系共振瑞利散射光谱研究及其分析应用
合成了巯基乙氨为稳定剂的CdTe量子点(QDs)。利用吸收光谱和共振瑞利散射光谱(RRS)研究了CdTe QDs与葡聚糖硫酸钠(DSS)的相互作用。CdTe QDs与DSS的吸收光谱在反应前后发生了明显的变化,这说明CdTe QDs与DSS之间存在着强烈的相互作用。在pH 5.0的缓冲溶液中,CdTe QDs-DSS体系的结合前后RRS变化显著。据此推测DSS通过静电引力和疏水作用力吸附到CdTe QDs表面形成了结合产物,导致了CdTe QDs-DSS体系的RRS显著增强。研究了最佳的反应条件和影响因素,同时研究了该方法的选择性。应用于血样和尿样的测定,结果令人满意。
Quantum dots (QDs), which are semiconductor nanoparticles that have all three dimensions confined to the 1-10 nm length scale, have attracted considerable attention because of their unique luminescent properties. QDs have been applied to the detection of biomacromolecules, metal ions, pharmaceuticals, and so on. Therefore, it is significant for further developing new analytical application of QDs.
Resonance Rayleigh scattering (RRS) technology was developed during 1990s. As a newly analytical technique, it has been received much attention because of its sensitivity and simplicity Therefore, this technique has been increasingly applied to the determination of macromolecules such as inorganic ions, organic compounds, nucleic acids, proteins, cationic surfactant, and some drugs. In recent years, this technique was applied to study nanometer particle. However, QDs were used as probes for the determination of polysaccharides by resonance Rayleigh scattering has few reports.
This paper contained two parts. The first part was the literature review and it includse the preparation and application of quantum dots, the development of resonance Rayleigh scattering technology and the introduction of polysaccharides in common analysis. The second part was research paper. In this paper, quantum dots were used as probes for the determination of two proteoglycans by resonance Rayleigh scattering method.
Determination of Dermatan Sulfate:
1. Resonance Rayleigh scattering method for the determination of Dermatan
Sulfate with CdSe quantum dots as probes
The CdSe quantum dots capped by 2-mercaptoethylamine hydrochloride with an average diameter of 2 rm were synthesized in water solution. It could react with dermatan sulfate (DS) and form the large aggregates by virtue of electrostatic attraction and the hydrophobic force, which induced a great enhancement of RRS. In a certain ranges, the enhancements of scattering intensity (ΔI) were directly proportional to the concentration of DS. This method revealed good sensitiviy, and the detection limit (38) was 1.3 ng mL-1. The optimum conditions of this reaction system and the influencing factors were investigated. In addition, ultraviolet absorption spectra and transmission electron microscope were employed to discuss the reaction mechanism of RRS enhancement. A novel method for the determination of trace DS using CdSe quantum dots probes was developed.
2. Resonance Rayleigh scattering spectra of CdTe QDs-DS system and its analytical application
The CdTe quantum dots capped by 2-mercaptoethylamine hydrochloride was synthesized in water solution. CdTe QDs can react with dermatan sulfate and form an ion association complex. The linear relactionship between the relative intensity ofΔIRRS and concentration of DS was in the range of 0.2-0.6μg mL-1. A sensitive, simple, and selective method for the determination of DS with CdTe QDs as probe by resonance Rayleigh scattering technique was developed. The optimum conditions and the influence factors were investigated. Moreover, the composition of ion-association complexes and the reaction mechanism were discussed. The method was applied to the determination of DS in human blood with satisfactory results.
Determination of Dextran Sulfate Sodium:
1. Resonance Rayleigh scattering method for the determination of dermatan sulfate sodium with water solubility CdSe quantum dots as probe
In acetic acid-sodium acetate buffer medium, the surface of QDs was assembled by 2-mercaptoethylamine hydrochloride which showed positive charge. The CdSe QDs could react with dextran sulfate sodium (DSS) and form the large aggregates by virtue of electrostatic attraction and the hydrophobic force, which leads to the remarkable enhancement of RRS. The maximum scattering peak was located at 343 nm. A sensitive and simple method for the determination of DSS with CdSe QDs as probes by resonance Rayleigh scattering technique was developed. The optimum conditions and influence factors of the reactions were investigated. It has been applied to the determination of DSS in human blood and urine samples and this method exhibited satisfactory results.
2. Resonance Rayleigh scattering spectra of CdTe QDs-DSS system and its analytical application
2-Mercaptoethylamine hydrochloride capped CdTe quantum dots with the diameter of 2-3 nm were synthesized in aqueous solution. At pH 5.0, CdTe QDs could react with DSS and form the large aggregates by electrostatic attraction and the hydrophobic force, which resulted in a great enhancement of resonance Rayleigh scattering. In this paper, the relationship between ultraviolet-visible absorption and resonance Rayleigh scattering was studied. The affecting factors and characteristics of RRS for the interaction of CdTe QDs with DSS were also investigated and a sensitive, simple and fast method for the determination of trace amounts of DSS using the RRS method was developed. It has been applied to the determination of DSS in human blood and urine samples with satisfactory results.
引文
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