单层配体保护的金纳米粒子合成及识别作用研究
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摘要
第一章：简要介绍了纳米材料的分类、特性及制备,并对金属纳米粒子尤其是金纳米粒子的光学性质及其分析应用进行了综述。
第二章：合成了β-环糊精衍生物全碘代β-环糊精和全巯基β-环糊精,以全巯基β-环糊精为稳定剂,用NaBH4还原HAuCl4·3H2O合成了全巯基β-环糊精单层保护的金纳米粒子(Per-thiol-β-CD-Au NPs),表面的全巯基β-环糊精既作为配体将金纳米保护起来,防止其聚集,使其粒径变小,确保其稳定性,又通过其外腔亲水使金纳米粒子易溶于水。结合紫外可见吸收光谱、FTIR,核磁共振氢谱、透射电子显微镜、质谱及热重分析对金纳米粒子进行了表征,由实验结果得知该金纳米的粒径大小约为2.7±0.8 nm,并估算出β-环糊精修饰的金纳米粒子中所含的平均金原子数和配体数分别为147和10个。
第三章：采用荧光光谱法研究了水溶性β-环糊精修饰的金纳米(多个结合位点超分子主体化合物)对L-酪氨酸的识别作用。L-酪氨酸本身在305nm处有一特征荧光发射峰,与β-环糊精修饰的金纳米作用后其荧光发生明显的猝灭。研究结果发现,在β-环糊精修饰的金纳米存在下,L-酪氨酸荧光强度的减小和其浓度呈线性关系。因此,基于此建立了一种用于测定L-酪氨酸的灵敏的荧光法,线性范围为0.02-1.5μM,检测限为1.6 nM, RSD为0.3%；并考察了温度的影响,以便更好的理解该体系猝灭原理和相应的结合作用力。研究结果表明该猝灭为静态猝灭且相应结合作用力以范德华力和疏水作用力为主。该方法已成功应用于实际样品复合氨基酸注射液中L-酪氨酸的定量测定,回收率在98.6～103.1%之间,取得了较满意的结果。
第四章：以N-乙酰基-L-半胱氨酸(NAC)为配体,用NaBH4还原,制备了Au/NAC摩尔比为1：3的NAC修饰的金纳米粒子,合成的金纳米粒子易溶于水,且溶液颜色呈棕色。应用红外光谱和紫外吸收光谱对其进行了表征,考察了金纳米粒子的荧光光谱特性。实验结果表明金纳米粒子的紫外吸收光谱范围宽,荧光最大发射为730 nm,处于近红外区；而且金纳米粒子的荧光光谱受pH的影响较小。我们利用荧光光谱法和紫外吸收光谱法研究了重金属离子(铜离子、银离子、汞离子和铅离子)对稳定的N-乙酰基-L-半胱氨酸修饰的水溶性金纳米粒子(NAC-Au NPs)荧光光谱的影响,研究结果表明重金属离子使NAC-Au NPs的近红外荧光发生了不同程度的猝灭；在此基础上进一步研究了pH对NAC-Au NPs荧光猝灭的影响。结合荧光光谱和紫外吸收光谱以及荧光寿命测量,我们提出了重金属离子对NAC修饰的金纳米粒子的可能猝灭机理。另研究发现在最佳条件下,加入EDTA可对共存金属离子进行掩蔽,但对汞离子掩蔽作用较小,会产生干扰。基于此,考虑可用于检测无汞离子的样品测定中,因此,建立了一种检测银离子的方法,线性范围为0-25μM,检测限为30nM,RSD为O.7%,采用标准加入法将其应用于矿泉水和自来水中银离子的测定,回收率在98～105%之间。这为水质检测领域提供了一定的理论依据。
Chapter 1:The classification, characteristics and synthesis of nanomaterials are depicted in brief. The optical properties and analytical application of noble metal nanoparticles, particularly gold nanoparticles, are reviewed in detail.
Chapter 2:Firstly, per-iodo- and per-thiol-β-cyclodextrin, that is, derivatives ofβ-cyclodextrin, were synthesized. Then, gold nanoparticles monolayer protected with per-thiol-β-cyclodextrin as a ligand or stable substrate was prepared, in which HAuCl4-3H2O was reduced by sodium triacetoxyborohydride. The nanoparticles were characterized by means of UV-visible absorption, FTIR, HNMR, Transmission Electron Microscope, Mass Spectroscopy and Thermogravimetric Analysis. Results indicated that the average size of P-cyclodextrin-modified gold nanoparticles is ca.2.7±0.8 run and numbers of gold atom and ligand were calculated to be 147 and 10, respectively.
Chapter 3:The interaction of water-solubleβ-cyclodextrin functionalized gold nanoparticles (β-CD-Au NPs) with L-tyrosine was studied by fluorescence spectroscopy. P-CD-Au NPs as an effective supramolecular host molecule in aqueous solution was found to remarkably quench fluorescence of L-tyrosine at the emission of 305 nm. Owing to the formation of the L-tyrosine-β-CD-Au NPs complex, static quenching involved resulted in the fluorescence quenching. Apparent binding constants and corresponding thermodynamic parameters at different temperatures were calculated by Stern-volmer equation and thermodynamic formula, respectively. Results indicated that hydrophobic interaction and Vader waals forces could be in dominant position in the formation of the complex. Under the optimum conditions, the concentration of L-tyrosine is proportional to decrease of the fluorescence intensity in the range from 0.02 to 1.5μM with the corresponding detection limit (S/N=3) of 1.6 nM and relative standard deviation is 0.3%. The developed method has been successfully employed to the quantitative determination of L-tyrosine in compound amino acid injection, and recovery ranges from 98.6 to 103.1%.
Chapter 4:N-Acetyl-L-Cysteine modified gold nanoaprticles with mole ratio from Au/NAC at 1:3 was synthesized, in which HAuCl4·3H2O was reduced by sodium triacetoxyborohydride. The as-prepared gold nanoparticles were soluble and the aqueous solution showed brown. They were chacterized by FTIR and UV-Vis spectroscopy. The fluorescence spectra of gold nanoparticles were also investigated. Experimental results indicated that UV-visible absorption spectra of NAC-Au NPs ranged from ultraviolet to visible regions, fluorescence maximum emission at 730 nm existed in the near IR region and the effect of pH on fluorescence spectra was little. In addition, interaction of water soluble NAC-Au NPs with metal ions containing copper ion, silver ion, mecury ion and lead ion was studied by fluorescence and UV-Vis spectroscopy. The presence of metal ions caused queching to different extent of NIR fluorescence of NAC-Au NPs and effect of pH was investigated, In combination of fluorescence and UV-Vis as well as time-resolved fluorescence measurement, the probably quenching mechanisms from different metal ions were proposed. The experimental results indicated that other coexistent metal ions apart from mercury ion were concealed in the presence of EDTA. Considering the application in the detection of real samples without mercury ions, the detection method of silver ion was developed. The linear range was 0-25μM, and detection limit was 30 nM. The method has been employed to the quantitative determination of silver ion in mianeral water and tap water with recovery in the range from 98 to 105%.
第二章：合成了β-环糊精衍生物全碘代β-环糊精和全巯基β-环糊精,以全巯基β-环糊精为稳定剂,用NaBH4还原HAuCl4·3H2O合成了全巯基β-环糊精单层保护的金纳米粒子(Per-thiol-β-CD-Au NPs),表面的全巯基β-环糊精既作为配体将金纳米保护起来,防止其聚集,使其粒径变小,确保其稳定性,又通过其外腔亲水使金纳米粒子易溶于水。结合紫外可见吸收光谱、FTIR,核磁共振氢谱、透射电子显微镜、质谱及热重分析对金纳米粒子进行了表征,由实验结果得知该金纳米的粒径大小约为2.7±0.8 nm,并估算出β-环糊精修饰的金纳米粒子中所含的平均金原子数和配体数分别为147和10个。
第三章：采用荧光光谱法研究了水溶性β-环糊精修饰的金纳米(多个结合位点超分子主体化合物)对L-酪氨酸的识别作用。L-酪氨酸本身在305nm处有一特征荧光发射峰,与β-环糊精修饰的金纳米作用后其荧光发生明显的猝灭。研究结果发现,在β-环糊精修饰的金纳米存在下,L-酪氨酸荧光强度的减小和其浓度呈线性关系。因此,基于此建立了一种用于测定L-酪氨酸的灵敏的荧光法,线性范围为0.02-1.5μM,检测限为1.6 nM, RSD为0.3%；并考察了温度的影响,以便更好的理解该体系猝灭原理和相应的结合作用力。研究结果表明该猝灭为静态猝灭且相应结合作用力以范德华力和疏水作用力为主。该方法已成功应用于实际样品复合氨基酸注射液中L-酪氨酸的定量测定,回收率在98.6～103.1%之间,取得了较满意的结果。
第四章：以N-乙酰基-L-半胱氨酸(NAC)为配体,用NaBH4还原,制备了Au/NAC摩尔比为1：3的NAC修饰的金纳米粒子,合成的金纳米粒子易溶于水,且溶液颜色呈棕色。应用红外光谱和紫外吸收光谱对其进行了表征,考察了金纳米粒子的荧光光谱特性。实验结果表明金纳米粒子的紫外吸收光谱范围宽,荧光最大发射为730 nm,处于近红外区；而且金纳米粒子的荧光光谱受pH的影响较小。我们利用荧光光谱法和紫外吸收光谱法研究了重金属离子(铜离子、银离子、汞离子和铅离子)对稳定的N-乙酰基-L-半胱氨酸修饰的水溶性金纳米粒子(NAC-Au NPs)荧光光谱的影响,研究结果表明重金属离子使NAC-Au NPs的近红外荧光发生了不同程度的猝灭；在此基础上进一步研究了pH对NAC-Au NPs荧光猝灭的影响。结合荧光光谱和紫外吸收光谱以及荧光寿命测量,我们提出了重金属离子对NAC修饰的金纳米粒子的可能猝灭机理。另研究发现在最佳条件下,加入EDTA可对共存金属离子进行掩蔽,但对汞离子掩蔽作用较小,会产生干扰。基于此,考虑可用于检测无汞离子的样品测定中,因此,建立了一种检测银离子的方法,线性范围为0-25μM,检测限为30nM,RSD为O.7%,采用标准加入法将其应用于矿泉水和自来水中银离子的测定,回收率在98～105%之间。这为水质检测领域提供了一定的理论依据。
Chapter 1:The classification, characteristics and synthesis of nanomaterials are depicted in brief. The optical properties and analytical application of noble metal nanoparticles, particularly gold nanoparticles, are reviewed in detail.
Chapter 2:Firstly, per-iodo- and per-thiol-β-cyclodextrin, that is, derivatives ofβ-cyclodextrin, were synthesized. Then, gold nanoparticles monolayer protected with per-thiol-β-cyclodextrin as a ligand or stable substrate was prepared, in which HAuCl4-3H2O was reduced by sodium triacetoxyborohydride. The nanoparticles were characterized by means of UV-visible absorption, FTIR, HNMR, Transmission Electron Microscope, Mass Spectroscopy and Thermogravimetric Analysis. Results indicated that the average size of P-cyclodextrin-modified gold nanoparticles is ca.2.7±0.8 run and numbers of gold atom and ligand were calculated to be 147 and 10, respectively.
Chapter 3:The interaction of water-solubleβ-cyclodextrin functionalized gold nanoparticles (β-CD-Au NPs) with L-tyrosine was studied by fluorescence spectroscopy. P-CD-Au NPs as an effective supramolecular host molecule in aqueous solution was found to remarkably quench fluorescence of L-tyrosine at the emission of 305 nm. Owing to the formation of the L-tyrosine-β-CD-Au NPs complex, static quenching involved resulted in the fluorescence quenching. Apparent binding constants and corresponding thermodynamic parameters at different temperatures were calculated by Stern-volmer equation and thermodynamic formula, respectively. Results indicated that hydrophobic interaction and Vader waals forces could be in dominant position in the formation of the complex. Under the optimum conditions, the concentration of L-tyrosine is proportional to decrease of the fluorescence intensity in the range from 0.02 to 1.5μM with the corresponding detection limit (S/N=3) of 1.6 nM and relative standard deviation is 0.3%. The developed method has been successfully employed to the quantitative determination of L-tyrosine in compound amino acid injection, and recovery ranges from 98.6 to 103.1%.
Chapter 4:N-Acetyl-L-Cysteine modified gold nanoaprticles with mole ratio from Au/NAC at 1:3 was synthesized, in which HAuCl4·3H2O was reduced by sodium triacetoxyborohydride. The as-prepared gold nanoparticles were soluble and the aqueous solution showed brown. They were chacterized by FTIR and UV-Vis spectroscopy. The fluorescence spectra of gold nanoparticles were also investigated. Experimental results indicated that UV-visible absorption spectra of NAC-Au NPs ranged from ultraviolet to visible regions, fluorescence maximum emission at 730 nm existed in the near IR region and the effect of pH on fluorescence spectra was little. In addition, interaction of water soluble NAC-Au NPs with metal ions containing copper ion, silver ion, mecury ion and lead ion was studied by fluorescence and UV-Vis spectroscopy. The presence of metal ions caused queching to different extent of NIR fluorescence of NAC-Au NPs and effect of pH was investigated, In combination of fluorescence and UV-Vis as well as time-resolved fluorescence measurement, the probably quenching mechanisms from different metal ions were proposed. The experimental results indicated that other coexistent metal ions apart from mercury ion were concealed in the presence of EDTA. Considering the application in the detection of real samples without mercury ions, the detection method of silver ion was developed. The linear range was 0-25μM, and detection limit was 30 nM. The method has been employed to the quantitative determination of silver ion in mianeral water and tap water with recovery in the range from 98 to 105%.
引文
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