金、碳纳米功能材料的制备及其色度和荧光传感方法研究
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摘要
本文以金、碳纳米材料为研究基础,以重要生物活性组分为研究对象,利用金纳米粒子(Gold nanoparticles, AuNPs)具有特殊的表面等离子体共振吸收、荧光金纳米簇(Gold nanoclusters, AuNCs)发射光波长随团簇尺寸可调、碳纳米材料(Carbon nanomaterials, CNMs)在能量转移特别是作为能量受体方面表现出来的优越性质,在合成制备的基础上,就基于金、碳纳米功能材料的色度和荧光传感方法开展研究,主要研究内容如下：
1.众所周知,ssDNA在AuNPs表面的选择性吸附能够稳定AuNPs,阻止高盐浓度诱导AuNPs的聚集,我们的研究表明,在精胺存在下,即使没有盐的额外加入,也能导致任意序列ssDNA包被的AuNPs发生非交联聚集。其机制在于精胺作为多价反离子,起到电荷屏蔽和“离子桥”的双重作用。就我们目前所知,由于阳离子多胺既没有吸收,也没有荧光等本征性光学特性,在发展关于阳离子多胺的光学分析方法方面,之前很少有成功的案例。因此,与传统的色谱和毛细管电泳相比,本文建立的方法将为生物体液和发酵食品中精胺的特异性检测提供一种简单方便的新选择。
2.碘离子作为一种生物重要活性阴离子,发展其简单、灵敏和特异性的检测方法仍然是一项有价值且富有挑战性的工作。在本文中,我们发现碘离子能够诱导组氨酸介导合成的荧光AuNCs的融合和各向异性生长,同时伴随AuNCs荧光的猝灭和裸眼可见溶液颜色的变化,据此建立了一种新的色度和荧光传感策略用于碘离子的特异性检测。与目前文献报道的方法相比,该方法具有简单和可视化检测的优点。其猝灭的荧光强度和增强表面等离子体共振吸收与碘离子浓度,分别在0.8～60和1.2～50μM具有线性关系,检测限(36)分别为118和215nM.
3.发现盐酸四环素能够在Nafion膜上产生非凡吸附,进一步研究表明其吸附是与浓度相关并由扩散控制的一级动力学过程,且是多层吸附并遵循BET吸附等温线。进一步,我们发现吸附在Nafion膜上的盐酸四环素,在合适浓度的氯金酸溶液中,能够介导金纳米晶体在Nafion膜上的生长,初步试验表明金纳米晶体的大小和形态可以通过改变氯金酸溶液的浓度进行简单调控。我们目前的初步试验结果,为金属纳米材料Nafion复合膜的制备提供了新的思路,相信这种金纳米材料Nafion聚合物复合膜在化学生物传感器设计和材料科学领域将具有潜在应用价值。
4.碳纳米管(Carbon nanotubes, CNTs)能够有效猝灭吸附到其表面荧光团的荧光,且ssDNA能够通过非共价相互作用缠绕到CNTs的表面。因此,荧光标记的ssDNA与碳纳米管能够形成自组装能量转移猝灭体系,基于采用核酸酶破坏该组装体的形成,可以观察到荧光恢复。基于这种机制,我们建立了一种荧光传感策略用于核酸酶活性的检测,其初始切割反应速率与核酸酶S1浓度在0.6～8.0U m1-1范围内具有线性关系,其检测限可达0.08U m1-1。进一步,我们以焦磷酸为例,用该方法去评价其对核酸酶活性的抑制效应,发现在0.2-1.4mM范围内,随着焦磷酸浓度的增加,荧光恢复的程度逐步减小,说明核酸酶对荧光标记的ssDNA切割反应受到了抑制,因此,发展的这种荧光核酸酶活性分析策略也可用于其潜在抑制剂的筛选。
5.分别比较研究了不同卟啉分子在GO和RGO上的组装,发现阳离子卟啉分子均能通过静电和π-π堆积作用自组装到两种石墨烯的表面,但由于卟啉分子在两种石墨烯上展平程度的差异,导致组装到RGO表面的卟啉分子出现更加明显的荧光猝灭和Soret吸收带的红移。随后,有趣的发现在于,阳离子卟啉与GO形成的复合物,能够易化和促进Fe(Ⅲ)螯合进入卟啉环,而阳离子卟啉与RGO形成的复合物却不能,原因在于与RGO相比,GO表面存在多种含氧功能基,可能起到辅助配位的作用。更有意思的是,Fe(Ⅲ)螯合进入卟啉环能够高效阻断从激态卟啉到GO的电子转移过程,从而导致卟啉分子荧光的恢复。据此,我们用阳离子卟啉与GO自组装形成的纳米杂交体作为光学探针,建立了Fe(Ⅲ)的荧光Turn-on传感方法,该方法对Fe(Ⅲ)展现出快速、灵敏的响应及高的选择性。
上述结果对深入认识两类纳米材料的功能性质,以进一步拓展其在纳米分析化学领域中的应用,具有重要的意义。
The various research results upon the nanomaterials reveal that they can open many good opportunities in an extremely multidisciplinary environment for promoting the rapid developments of different research fields. For example, gold nanoparticles have wide-range use as a fertile ground for analytical purpose due to the character of strong surface plasmon resonance and ease of observation by naked eyes, and gold nanoclusters have molecule-like characteristics which make it give rise to unique and size dependent fluorescent properties. Additionally, it is well known that carbon nanotubes (CNTs) and graphene can nonconvalently interact with soft single-stranded DNA (ssDNA) by means of π-stacking interactions between nucleotide bases and carbon nanomaterials described above, and can also act collectively as quenchers for the fluorophores owing to energy-transfer and electron-transfer processes. Therefore, considering their promising optical charaters, in this conribution we attempt to develope the colorimetric and fluorometric sensing for the biologically important molecules based on the gold and carbon nanomaterials. The main contains are listed as follows:
1. The selective adsorption of single-stranded oligonucleotides (ssDNA) on gold nanoparticles (AuNPs) is well known for stabilizing the AuNPs against aggregation even at high salt concentrations. Our investigation shows that the non-crosslinking aggregation of arbitrary ssDNA-capped AuNPs occurs due to their interaction with the cationic polyamine, spermine (Spm), even without any addition of NaCl. The non-crosslinking aggregation mechanism is that the Spm, served as multivalent counterions, plays the dual roles of charge shielding and ion bridging among the ssDNA-capped AuNPs, which jointly result in the aggregation of the ssDNA-capped AuNPs. Therefore, a sensitive and highly selective colorimetric method for the detection of Spm was developed. To the best of our knowledge, it is the first successful case as to the efforts towards the development of optical assays for cationic polyamine, showing neither natural UV absorption nor fluorescence. Compared with the traditional methods of chromatography and capillary electrophoresis, the approach described here would provide a convenient alternative and new train of thought for the specific detection of Spm in both biological fluid and fermented products.
2. Iodide, as a biologically important anion, it remains a worthwhile yet challenging undertaking to find a sensitive and specific approach to provide a technically simple iodide detection. In this article, it was found that no other ions than iodide-induced anisotropic growth of AuNCs originated from a small molecule, histidine-mediated synthesis of AuNCs, were observed. Simultaneously, it is accompanied by the fluorescence quenching of AuNCs and the naked-eye visible color change. Therefore, a new colorimetric and fluorometric sensing strategy was developed for the iodide-specific detection. Compared with currently reported methods, the present one displays the advantages of the visual detection and simplicity. The quenched fluorescence and enhanced surface plasmon resonance absorbance were found to be proportional to the iodide concentration over the range of0.8-60and1.2-50μM with a detection limit (3a) of118nM and215nM, respectively.
3. Tetracycline hydrochloride (Tc) was found to be adsorbed well into a Nafion (Nf) film. The kenetic analysis suggested that the adsorption of Tc into the Nf film is controlled by its diffusion in the Nf film with a diffusion coefficient of D=8.5x10-9cm2s-1The adsorption isothermal was analyzed by a Brunauer-Emmett-Teller (BET) equation suggertion multilayer adsorption of Tc into the film. Further, the growth of gold nanocrystals could be observed if the Nf film adsorbed the Tc was dipped into the HAuCl4solution, and the preliminary experimental results demonstrate that the shape of gold nanocrystals can be controlled by simply changing the concentration of HAuCl4solution. The research results are significant and the approach described here would provide new train of thought to develope nanocomposites consisting of metal nanoparticles embedded in a polymer matrix, considering that they could make new materials with novel properties originating from the combination of the inorganic components and the polymer and have promising applications in the field of optical sensing.
4. Carbon nanotubes (CNTs) can efficiently quench the fluorescence of the adsorbed fluorophores and nonconvalently interact with soft single-stranded DNA (ssDNA). Upon disruption of CNTs-fluorescent oligonucleotides hybrid by nuclease S1, fluorescence turn-on was observed. Using this strategy, a platform based on fluorescence signal for monitoring the activity of nuclease with advantages of high sensitivity and commonality was established, and a linear relationship between initial cleavage reaction rate and nuclease S1concentration is found in the range of0.6-8.0U ml-1with a detection limit of0.08U ml-1. Furthermore, by taking pyrophosphate as an example, we use the assay to evaluate the prohibition effect on nuclease, and the extent of fluorescence recovery decreased linearly with increasing the concentration of pyrophosphate in the range of0.2-1.4mM, implying that the cleavage reaction by nuclease S1was prohibited, and therefore this fluorescence assay can also be conveniently utilized for inhibitor screening of nuclease.
5. A comparative reaserch of the assembly of different porphyrin molecules on graphene oxide (GO) and reduced graphene oxide (RGO) was carried out, respectively. Despite the cationic porphyrin molecules can be assembled onto the surfaces of graphene sheets, including GO and RGO, to form complexes through electrostatic and π-π stacking interactions, the more obvious fluorescence quenching and the larger red-shift of the Soret band of porphyrin molecule in RGO-bound states were observed than those in GO-bound states, due to the differenc of molecular flattening in degree. Further, more interesting finding was that the complexes formed between cationic porphyrin and GO, rather than RGO sheets, can facilitate the incorporation of iron (Ⅲ) ions into the porphyrin moieties, due to the presence of the oxygen-contained groups at the basal plane of GO sheets served as auxiliary coordination units, which can high-efficiently obstruct the electron transfer from excited porphyrin to GO sheets and result in the occurrence of fluorescence restoration. Thus, a fluorescence sensing platform has been developed for iron (Ⅲ) ions detection in this contribution by using the porphyrin/GO nanohybrids as an optical probe, and our present one exhibited rapid and sensitive responses and high selectivity toward iron (Ⅲ) ions.
In summary, we suppose that the research results describes above would be beneficial for us to understand deeply their characters of the two kinds of nanomaterials, and it is very important for us to enlarge their applications in nanomaterial-based analytical chemistry.
1.众所周知,ssDNA在AuNPs表面的选择性吸附能够稳定AuNPs,阻止高盐浓度诱导AuNPs的聚集,我们的研究表明,在精胺存在下,即使没有盐的额外加入,也能导致任意序列ssDNA包被的AuNPs发生非交联聚集。其机制在于精胺作为多价反离子,起到电荷屏蔽和“离子桥”的双重作用。就我们目前所知,由于阳离子多胺既没有吸收,也没有荧光等本征性光学特性,在发展关于阳离子多胺的光学分析方法方面,之前很少有成功的案例。因此,与传统的色谱和毛细管电泳相比,本文建立的方法将为生物体液和发酵食品中精胺的特异性检测提供一种简单方便的新选择。
2.碘离子作为一种生物重要活性阴离子,发展其简单、灵敏和特异性的检测方法仍然是一项有价值且富有挑战性的工作。在本文中,我们发现碘离子能够诱导组氨酸介导合成的荧光AuNCs的融合和各向异性生长,同时伴随AuNCs荧光的猝灭和裸眼可见溶液颜色的变化,据此建立了一种新的色度和荧光传感策略用于碘离子的特异性检测。与目前文献报道的方法相比,该方法具有简单和可视化检测的优点。其猝灭的荧光强度和增强表面等离子体共振吸收与碘离子浓度,分别在0.8～60和1.2～50μM具有线性关系,检测限(36)分别为118和215nM.
3.发现盐酸四环素能够在Nafion膜上产生非凡吸附,进一步研究表明其吸附是与浓度相关并由扩散控制的一级动力学过程,且是多层吸附并遵循BET吸附等温线。进一步,我们发现吸附在Nafion膜上的盐酸四环素,在合适浓度的氯金酸溶液中,能够介导金纳米晶体在Nafion膜上的生长,初步试验表明金纳米晶体的大小和形态可以通过改变氯金酸溶液的浓度进行简单调控。我们目前的初步试验结果,为金属纳米材料Nafion复合膜的制备提供了新的思路,相信这种金纳米材料Nafion聚合物复合膜在化学生物传感器设计和材料科学领域将具有潜在应用价值。
4.碳纳米管(Carbon nanotubes, CNTs)能够有效猝灭吸附到其表面荧光团的荧光,且ssDNA能够通过非共价相互作用缠绕到CNTs的表面。因此,荧光标记的ssDNA与碳纳米管能够形成自组装能量转移猝灭体系,基于采用核酸酶破坏该组装体的形成,可以观察到荧光恢复。基于这种机制,我们建立了一种荧光传感策略用于核酸酶活性的检测,其初始切割反应速率与核酸酶S1浓度在0.6～8.0U m1-1范围内具有线性关系,其检测限可达0.08U m1-1。进一步,我们以焦磷酸为例,用该方法去评价其对核酸酶活性的抑制效应,发现在0.2-1.4mM范围内,随着焦磷酸浓度的增加,荧光恢复的程度逐步减小,说明核酸酶对荧光标记的ssDNA切割反应受到了抑制,因此,发展的这种荧光核酸酶活性分析策略也可用于其潜在抑制剂的筛选。
5.分别比较研究了不同卟啉分子在GO和RGO上的组装,发现阳离子卟啉分子均能通过静电和π-π堆积作用自组装到两种石墨烯的表面,但由于卟啉分子在两种石墨烯上展平程度的差异,导致组装到RGO表面的卟啉分子出现更加明显的荧光猝灭和Soret吸收带的红移。随后,有趣的发现在于,阳离子卟啉与GO形成的复合物,能够易化和促进Fe(Ⅲ)螯合进入卟啉环,而阳离子卟啉与RGO形成的复合物却不能,原因在于与RGO相比,GO表面存在多种含氧功能基,可能起到辅助配位的作用。更有意思的是,Fe(Ⅲ)螯合进入卟啉环能够高效阻断从激态卟啉到GO的电子转移过程,从而导致卟啉分子荧光的恢复。据此,我们用阳离子卟啉与GO自组装形成的纳米杂交体作为光学探针,建立了Fe(Ⅲ)的荧光Turn-on传感方法,该方法对Fe(Ⅲ)展现出快速、灵敏的响应及高的选择性。
上述结果对深入认识两类纳米材料的功能性质,以进一步拓展其在纳米分析化学领域中的应用,具有重要的意义。
The various research results upon the nanomaterials reveal that they can open many good opportunities in an extremely multidisciplinary environment for promoting the rapid developments of different research fields. For example, gold nanoparticles have wide-range use as a fertile ground for analytical purpose due to the character of strong surface plasmon resonance and ease of observation by naked eyes, and gold nanoclusters have molecule-like characteristics which make it give rise to unique and size dependent fluorescent properties. Additionally, it is well known that carbon nanotubes (CNTs) and graphene can nonconvalently interact with soft single-stranded DNA (ssDNA) by means of π-stacking interactions between nucleotide bases and carbon nanomaterials described above, and can also act collectively as quenchers for the fluorophores owing to energy-transfer and electron-transfer processes. Therefore, considering their promising optical charaters, in this conribution we attempt to develope the colorimetric and fluorometric sensing for the biologically important molecules based on the gold and carbon nanomaterials. The main contains are listed as follows:
1. The selective adsorption of single-stranded oligonucleotides (ssDNA) on gold nanoparticles (AuNPs) is well known for stabilizing the AuNPs against aggregation even at high salt concentrations. Our investigation shows that the non-crosslinking aggregation of arbitrary ssDNA-capped AuNPs occurs due to their interaction with the cationic polyamine, spermine (Spm), even without any addition of NaCl. The non-crosslinking aggregation mechanism is that the Spm, served as multivalent counterions, plays the dual roles of charge shielding and ion bridging among the ssDNA-capped AuNPs, which jointly result in the aggregation of the ssDNA-capped AuNPs. Therefore, a sensitive and highly selective colorimetric method for the detection of Spm was developed. To the best of our knowledge, it is the first successful case as to the efforts towards the development of optical assays for cationic polyamine, showing neither natural UV absorption nor fluorescence. Compared with the traditional methods of chromatography and capillary electrophoresis, the approach described here would provide a convenient alternative and new train of thought for the specific detection of Spm in both biological fluid and fermented products.
2. Iodide, as a biologically important anion, it remains a worthwhile yet challenging undertaking to find a sensitive and specific approach to provide a technically simple iodide detection. In this article, it was found that no other ions than iodide-induced anisotropic growth of AuNCs originated from a small molecule, histidine-mediated synthesis of AuNCs, were observed. Simultaneously, it is accompanied by the fluorescence quenching of AuNCs and the naked-eye visible color change. Therefore, a new colorimetric and fluorometric sensing strategy was developed for the iodide-specific detection. Compared with currently reported methods, the present one displays the advantages of the visual detection and simplicity. The quenched fluorescence and enhanced surface plasmon resonance absorbance were found to be proportional to the iodide concentration over the range of0.8-60and1.2-50μM with a detection limit (3a) of118nM and215nM, respectively.
3. Tetracycline hydrochloride (Tc) was found to be adsorbed well into a Nafion (Nf) film. The kenetic analysis suggested that the adsorption of Tc into the Nf film is controlled by its diffusion in the Nf film with a diffusion coefficient of D=8.5x10-9cm2s-1The adsorption isothermal was analyzed by a Brunauer-Emmett-Teller (BET) equation suggertion multilayer adsorption of Tc into the film. Further, the growth of gold nanocrystals could be observed if the Nf film adsorbed the Tc was dipped into the HAuCl4solution, and the preliminary experimental results demonstrate that the shape of gold nanocrystals can be controlled by simply changing the concentration of HAuCl4solution. The research results are significant and the approach described here would provide new train of thought to develope nanocomposites consisting of metal nanoparticles embedded in a polymer matrix, considering that they could make new materials with novel properties originating from the combination of the inorganic components and the polymer and have promising applications in the field of optical sensing.
4. Carbon nanotubes (CNTs) can efficiently quench the fluorescence of the adsorbed fluorophores and nonconvalently interact with soft single-stranded DNA (ssDNA). Upon disruption of CNTs-fluorescent oligonucleotides hybrid by nuclease S1, fluorescence turn-on was observed. Using this strategy, a platform based on fluorescence signal for monitoring the activity of nuclease with advantages of high sensitivity and commonality was established, and a linear relationship between initial cleavage reaction rate and nuclease S1concentration is found in the range of0.6-8.0U ml-1with a detection limit of0.08U ml-1. Furthermore, by taking pyrophosphate as an example, we use the assay to evaluate the prohibition effect on nuclease, and the extent of fluorescence recovery decreased linearly with increasing the concentration of pyrophosphate in the range of0.2-1.4mM, implying that the cleavage reaction by nuclease S1was prohibited, and therefore this fluorescence assay can also be conveniently utilized for inhibitor screening of nuclease.
5. A comparative reaserch of the assembly of different porphyrin molecules on graphene oxide (GO) and reduced graphene oxide (RGO) was carried out, respectively. Despite the cationic porphyrin molecules can be assembled onto the surfaces of graphene sheets, including GO and RGO, to form complexes through electrostatic and π-π stacking interactions, the more obvious fluorescence quenching and the larger red-shift of the Soret band of porphyrin molecule in RGO-bound states were observed than those in GO-bound states, due to the differenc of molecular flattening in degree. Further, more interesting finding was that the complexes formed between cationic porphyrin and GO, rather than RGO sheets, can facilitate the incorporation of iron (Ⅲ) ions into the porphyrin moieties, due to the presence of the oxygen-contained groups at the basal plane of GO sheets served as auxiliary coordination units, which can high-efficiently obstruct the electron transfer from excited porphyrin to GO sheets and result in the occurrence of fluorescence restoration. Thus, a fluorescence sensing platform has been developed for iron (Ⅲ) ions detection in this contribution by using the porphyrin/GO nanohybrids as an optical probe, and our present one exhibited rapid and sensitive responses and high selectivity toward iron (Ⅲ) ions.
In summary, we suppose that the research results describes above would be beneficial for us to understand deeply their characters of the two kinds of nanomaterials, and it is very important for us to enlarge their applications in nanomaterial-based analytical chemistry.
引文
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