高铁酸盐、DNA传感器的绿色分析化学方法研究
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摘要
作为当今国际化学学科研究的前沿,绿色化学受到了广泛重视。绿色分析化学是绿色化学的重要组成部分,它是把绿色化学的原理应用在新的分析方法中,旨在减轻分析化学对环境的影响。今后绿色分析化学发展的主要方向将是建立无污染或少污染的分析化学方法,从而满足可持续发展的要求。本文着重介绍了高铁酸盐以及电化学DNA传感器的绿色分析化学方法研究,主要研究内容和结果如下:
(1)基于高铁酸盐可以氧化鲁米诺发光这一特性,建立了一种新型、快速、环境友好的在线监测高铁酸盐溶液稳定性的方法——流动注射–化学发光分析法。研究了温度、溶液pH值、添加剂对高铁酸盐溶液稳定性的影响。实验结果表明低温和高碱度是保持高铁酸盐溶液稳定的关键因素。硅酸钠和碘化钾作为添加剂有助于提高高铁酸盐的产率和稳定性;氯化钠对高铁酸盐溶液有加速分解的作用。
(2)实验建立了一种新型的快速处理阳极表面钝化膜的方法——草酸浸泡法用于提高高铁酸盐的产率。通过SEM和XPS分别对电极的表面的形貌和化学成分进行了分析。和阴极极化法相比,草酸浸泡法具有较高的阳极钝化膜去除效率,且去除后的电极表面呈现疏松的多孔结构,有利于提高高铁酸盐电化学合成产率。另外,实验采用流动注射-化学发光分析法研究了草酸浸泡处理后的铁阳极电解制备高铁酸盐溶液的稳定性和重现性。实验结果表明在低温下高铁酸盐溶液具有较好的稳定性,且6次重复电解所得的高铁酸盐溶液化学发光信号重现性较好,其相对标准偏差为2.41%。
(3)实验在低浓度NaOH条件下成功合成了高铁酸盐溶液。在0±1oC下,高铁酸盐溶液能在较长时间内保持相对稳定。基于此,实验选取鲁米诺为研究对象,验证了低浓度NaOH条件下电解合成的高铁酸盐溶液用于流动注射-化学发光技术的可行性。在优化条件下,鲁米诺的线性范围为5.0×10~(-9) ~ 1.0×10~(-6) mol/L;检测限为(S/N = 3) 2.5×10~(-9) mol/L;对3.0×10-7 mol/L的鲁米诺进行6次平行测定的相对标准偏差(RSD)为2.57%。
(4)研究了低浓度NaOH条件下电解制备高铁酸盐溶液直接用于流动注射-化学发光检测的新方法。基于无机离子、有机分子和生物分子对鲁米诺–高铁酸盐化学发光反应的增敏作用,该方法成功实现了对V(V)、Ca(II)、Mg(II)、间苯三酚和牛血红蛋白的分析测定。在优化实验条件下,V(V)、Ca(II)、Mg(II)、间苯三酚和牛血红蛋白的检测限(S/N = 3)分别为1.96×10~(-10) mol/L,1.25×10~(-9) mol/L,1.67×10~(-7) mol/L,1.00×10~(-8) mol/L,1.24×10~(-12) mol/L。对3.92×10~(-9) mol/L V(V),1.25×10~(-7) mol/L Ca(II),4.17×10~(-5) mol/L Mg(II),2.00×10~(-7) mol/L间苯三酚和3.10×10~(-9) mol/L牛血红蛋白进行6次测定的相对标准偏差分别为2.63%, 2.38%, 1.32%, 0.72 %和2.74%。
(5)研究了低浓度NaOH条件下电解制备高铁酸盐溶液直接用于毛细管电泳-化学发光检测的新方法。实验选取8-羟基喹啉为检测对象,在优化条件下,其的线性范围为5.0×10~(-10) ~ 5.0×10~(-8) mol/L (R~2 = 0.997);检测限为(S/N = 3) 5.0×10~(-10) mol/L;对2.5×10~(-9) mol/L的8-羟基喹啉进行5次平行测定的相对标准偏差(RSD)为1.43%。
(6)研制出了一种新型的磁性金电极,该电极具有诸多优良特性,比如具有良好的导电性,温和的磁场强度,耐用,操作简单,能快速将DNA标记的磁珠富集到电极表面。基于该电极的DNA电化学传感器成功用于DNA杂交的检测。首先,巯基修饰的探针DNA自组装到金磁纳米珠表面;目标DNA与捕获探针杂交,并选用亚甲基蓝为杂交指示剂;随后,捕获探针与目标DNA的双链复合物修饰的金磁纳米珠被富集到磁电极表面进行电化学检测。构造的传感器检测完全互补的目标DNA的线性范围在0.3 ~ 300 nM,检测限为0.1 nM。另外,该传感器还具有良好的选择性。
Green chemistry is a frontier area of chemistry, which has attracted abroad attention. Green analytical chemistry plays an important role in green chemistry. The goal of green analytical chemistry is to use analytical procedures that are safe to environmental. This may be achieved by developing new analytical methodologies that are more environmental friendly. The present study was therefore designed to research on the green analytical chemistry methods of ferrate(VI) and DNA biosensor. The detailed contents are described as follows.
(1) Based on the homogeneous chemiluminescence (CL) reaction of luminol oxidized by ferrate(VI), a novel, convenient, environmental friendly, on-line monitoring method was developed to investigate the stability of ferrate(VI) solution. The effects of temperature, pH, additives on the stability of ferrate(VI) solution were traced by flow injection–chemiluminescence (FI–CL) method. It was found that the lowe storage temperature and high pH were the key factors for the stability of ferrate(VI) solution. Na2SiO3 and KI could enhance the synthesis efficiency and the stability of ferrate(VI) solution; NaCl could catalyze the decomposition of ferrate(VI) solution.
(2) A convenient anodic pretreatment was established based on the possibility of dissolution of passivation using oxalic acid. Then, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) examinations were used to examine the structure and composition of anode surface, respectively. The high removal efficiency of passivation and the formation of porous structure on anode surface could be observed. These led to significant enhancement of the ferrate(VI) synthesis efficiency. The characteristics of ferrate(VI) solution such as stability and reproducibility were studied by FI–CL method based on the CL reaction of ferrate(VI)–luminol. The CL signal was stable for a week. The relative standard deviation (RSD) was 2.41% for six runs of electrolysis.
(3) It was found ferrate(VI) could be successfully electrogenerated in low-concentration NaOH solution for direct analytical application. The characteristic of ferrate(VI) solution is relatively stable at 0±1oC. Luminol chosen as samples was detected by FI–CL method. The detailed analysis and testing revealed that luminol showed a linear CL response in the concentration range of 5.0×10~(-9) ~ 1.0×10~(-6) mol/L (R~2 = 0.9964), with a detection limit (S/N = 3) of 2.5×10~(-9) mol/L. The RSD for 6 repeated measurements of 3.0×10~(-7) mol/L luminol was 2.57%.
(4) The possibility of direct analytical applications of ferrate(VI) solution, which was freshly electrogenerated in low-concentration NaOH electrolyte, was studied by a FI–CL system. It was found that some inorganic ions, organic molecule and biomolecule could enhance the chemiluminescence emission caused by ferrate(VI)–luminol reaction. V(V), Ca(II), Mg(II), phloroglucinol, and bovine hemoglobin (Hb) chosen as samples were successfully detected by this developed method. The analytical characteristics of the system for the analytes determination including linear ranges, correlation coefficients, limits of detection combined with FI analysis were studied. The limits of detection for V(V), Ca(II), Mg(II), phloroglucinol and Hb were 1.96×10~(-10), 1.25×10~(-9), 1.67×10~(-7), 1.00×10~(-8), and 1.24×10~(-12) mol/L, respectively. The RSD for six repeated measurements of 3.92×10~(-9), 1.25×10~(-7), 4.17×10~(-5), 2.00×10~(-7) and 3.10×10~(-9) mol/L of V(V), Ca(II), Mg(II), phloroglucinol and Hb were 2.63, 2.38, 1.32, 0.72 and 2.74% , respectively.
(5) A novel, eco-friendly, and stable homogeneous luminescent system for capillary zone electrophoresis (CE) detection was established, based on the CL flash caused by mixing ferrate(VI) with luminol solution. To test this CL system, 8-hydroxyquinoline (HQ) was chosen as a modal sample by CE detection. The detailed analysis and testing revealed that HQ showed a linear CL response in the concentration range of 5.0×10~(-10) ~ 5.0×10~(-8) mol/L (R~2 = 0.997), with a detection limit (S/N = 3) of 5.0×10~(-10) mol/L. For 2.5×10~(-9) mol/L HQ, when the repeated injections were performed with the same run of electrolyzed ferrate(VI), the RSD of peak height was 1.43% (n = 5).
(6) A simple method was proposed to prepare magnetic gold electrodes for DNA hybridization detection. The magnetic gold electrodes displayed several excellent performances such as good electrical conductivity, moderate magnetic field intensity, easy to capture and enrich probe-magnetic beads onto the electrode surface, robust and simple to operate. A thiolated capture probe DNA was self-assembled on gold magnetic nanobeads (GMNBs) by gold-sufur affinity. The probe DNA-GMNBs selectively hybridized with the target DNA. Methylene blue (Mb) was then used as the electrochemical intercalator to indicate DNA hybridization. The target DNA/probe DNA/GMNBs were enriched onto the electrode surface for detection. The peak current of Mb linearly decreased with the concentration of the complementary target DNA over a range from 0.3 to 300 nM with a detection limit of 0.1 nM. Furthermore, the selectivity of this biosensor for DNA hybridization was successfully examined.
(1)基于高铁酸盐可以氧化鲁米诺发光这一特性,建立了一种新型、快速、环境友好的在线监测高铁酸盐溶液稳定性的方法——流动注射–化学发光分析法。研究了温度、溶液pH值、添加剂对高铁酸盐溶液稳定性的影响。实验结果表明低温和高碱度是保持高铁酸盐溶液稳定的关键因素。硅酸钠和碘化钾作为添加剂有助于提高高铁酸盐的产率和稳定性;氯化钠对高铁酸盐溶液有加速分解的作用。
(2)实验建立了一种新型的快速处理阳极表面钝化膜的方法——草酸浸泡法用于提高高铁酸盐的产率。通过SEM和XPS分别对电极的表面的形貌和化学成分进行了分析。和阴极极化法相比,草酸浸泡法具有较高的阳极钝化膜去除效率,且去除后的电极表面呈现疏松的多孔结构,有利于提高高铁酸盐电化学合成产率。另外,实验采用流动注射-化学发光分析法研究了草酸浸泡处理后的铁阳极电解制备高铁酸盐溶液的稳定性和重现性。实验结果表明在低温下高铁酸盐溶液具有较好的稳定性,且6次重复电解所得的高铁酸盐溶液化学发光信号重现性较好,其相对标准偏差为2.41%。
(3)实验在低浓度NaOH条件下成功合成了高铁酸盐溶液。在0±1oC下,高铁酸盐溶液能在较长时间内保持相对稳定。基于此,实验选取鲁米诺为研究对象,验证了低浓度NaOH条件下电解合成的高铁酸盐溶液用于流动注射-化学发光技术的可行性。在优化条件下,鲁米诺的线性范围为5.0×10~(-9) ~ 1.0×10~(-6) mol/L;检测限为(S/N = 3) 2.5×10~(-9) mol/L;对3.0×10-7 mol/L的鲁米诺进行6次平行测定的相对标准偏差(RSD)为2.57%。
(4)研究了低浓度NaOH条件下电解制备高铁酸盐溶液直接用于流动注射-化学发光检测的新方法。基于无机离子、有机分子和生物分子对鲁米诺–高铁酸盐化学发光反应的增敏作用,该方法成功实现了对V(V)、Ca(II)、Mg(II)、间苯三酚和牛血红蛋白的分析测定。在优化实验条件下,V(V)、Ca(II)、Mg(II)、间苯三酚和牛血红蛋白的检测限(S/N = 3)分别为1.96×10~(-10) mol/L,1.25×10~(-9) mol/L,1.67×10~(-7) mol/L,1.00×10~(-8) mol/L,1.24×10~(-12) mol/L。对3.92×10~(-9) mol/L V(V),1.25×10~(-7) mol/L Ca(II),4.17×10~(-5) mol/L Mg(II),2.00×10~(-7) mol/L间苯三酚和3.10×10~(-9) mol/L牛血红蛋白进行6次测定的相对标准偏差分别为2.63%, 2.38%, 1.32%, 0.72 %和2.74%。
(5)研究了低浓度NaOH条件下电解制备高铁酸盐溶液直接用于毛细管电泳-化学发光检测的新方法。实验选取8-羟基喹啉为检测对象,在优化条件下,其的线性范围为5.0×10~(-10) ~ 5.0×10~(-8) mol/L (R~2 = 0.997);检测限为(S/N = 3) 5.0×10~(-10) mol/L;对2.5×10~(-9) mol/L的8-羟基喹啉进行5次平行测定的相对标准偏差(RSD)为1.43%。
(6)研制出了一种新型的磁性金电极,该电极具有诸多优良特性,比如具有良好的导电性,温和的磁场强度,耐用,操作简单,能快速将DNA标记的磁珠富集到电极表面。基于该电极的DNA电化学传感器成功用于DNA杂交的检测。首先,巯基修饰的探针DNA自组装到金磁纳米珠表面;目标DNA与捕获探针杂交,并选用亚甲基蓝为杂交指示剂;随后,捕获探针与目标DNA的双链复合物修饰的金磁纳米珠被富集到磁电极表面进行电化学检测。构造的传感器检测完全互补的目标DNA的线性范围在0.3 ~ 300 nM,检测限为0.1 nM。另外,该传感器还具有良好的选择性。
Green chemistry is a frontier area of chemistry, which has attracted abroad attention. Green analytical chemistry plays an important role in green chemistry. The goal of green analytical chemistry is to use analytical procedures that are safe to environmental. This may be achieved by developing new analytical methodologies that are more environmental friendly. The present study was therefore designed to research on the green analytical chemistry methods of ferrate(VI) and DNA biosensor. The detailed contents are described as follows.
(1) Based on the homogeneous chemiluminescence (CL) reaction of luminol oxidized by ferrate(VI), a novel, convenient, environmental friendly, on-line monitoring method was developed to investigate the stability of ferrate(VI) solution. The effects of temperature, pH, additives on the stability of ferrate(VI) solution were traced by flow injection–chemiluminescence (FI–CL) method. It was found that the lowe storage temperature and high pH were the key factors for the stability of ferrate(VI) solution. Na2SiO3 and KI could enhance the synthesis efficiency and the stability of ferrate(VI) solution; NaCl could catalyze the decomposition of ferrate(VI) solution.
(2) A convenient anodic pretreatment was established based on the possibility of dissolution of passivation using oxalic acid. Then, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) examinations were used to examine the structure and composition of anode surface, respectively. The high removal efficiency of passivation and the formation of porous structure on anode surface could be observed. These led to significant enhancement of the ferrate(VI) synthesis efficiency. The characteristics of ferrate(VI) solution such as stability and reproducibility were studied by FI–CL method based on the CL reaction of ferrate(VI)–luminol. The CL signal was stable for a week. The relative standard deviation (RSD) was 2.41% for six runs of electrolysis.
(3) It was found ferrate(VI) could be successfully electrogenerated in low-concentration NaOH solution for direct analytical application. The characteristic of ferrate(VI) solution is relatively stable at 0±1oC. Luminol chosen as samples was detected by FI–CL method. The detailed analysis and testing revealed that luminol showed a linear CL response in the concentration range of 5.0×10~(-9) ~ 1.0×10~(-6) mol/L (R~2 = 0.9964), with a detection limit (S/N = 3) of 2.5×10~(-9) mol/L. The RSD for 6 repeated measurements of 3.0×10~(-7) mol/L luminol was 2.57%.
(4) The possibility of direct analytical applications of ferrate(VI) solution, which was freshly electrogenerated in low-concentration NaOH electrolyte, was studied by a FI–CL system. It was found that some inorganic ions, organic molecule and biomolecule could enhance the chemiluminescence emission caused by ferrate(VI)–luminol reaction. V(V), Ca(II), Mg(II), phloroglucinol, and bovine hemoglobin (Hb) chosen as samples were successfully detected by this developed method. The analytical characteristics of the system for the analytes determination including linear ranges, correlation coefficients, limits of detection combined with FI analysis were studied. The limits of detection for V(V), Ca(II), Mg(II), phloroglucinol and Hb were 1.96×10~(-10), 1.25×10~(-9), 1.67×10~(-7), 1.00×10~(-8), and 1.24×10~(-12) mol/L, respectively. The RSD for six repeated measurements of 3.92×10~(-9), 1.25×10~(-7), 4.17×10~(-5), 2.00×10~(-7) and 3.10×10~(-9) mol/L of V(V), Ca(II), Mg(II), phloroglucinol and Hb were 2.63, 2.38, 1.32, 0.72 and 2.74% , respectively.
(5) A novel, eco-friendly, and stable homogeneous luminescent system for capillary zone electrophoresis (CE) detection was established, based on the CL flash caused by mixing ferrate(VI) with luminol solution. To test this CL system, 8-hydroxyquinoline (HQ) was chosen as a modal sample by CE detection. The detailed analysis and testing revealed that HQ showed a linear CL response in the concentration range of 5.0×10~(-10) ~ 5.0×10~(-8) mol/L (R~2 = 0.997), with a detection limit (S/N = 3) of 5.0×10~(-10) mol/L. For 2.5×10~(-9) mol/L HQ, when the repeated injections were performed with the same run of electrolyzed ferrate(VI), the RSD of peak height was 1.43% (n = 5).
(6) A simple method was proposed to prepare magnetic gold electrodes for DNA hybridization detection. The magnetic gold electrodes displayed several excellent performances such as good electrical conductivity, moderate magnetic field intensity, easy to capture and enrich probe-magnetic beads onto the electrode surface, robust and simple to operate. A thiolated capture probe DNA was self-assembled on gold magnetic nanobeads (GMNBs) by gold-sufur affinity. The probe DNA-GMNBs selectively hybridized with the target DNA. Methylene blue (Mb) was then used as the electrochemical intercalator to indicate DNA hybridization. The target DNA/probe DNA/GMNBs were enriched onto the electrode surface for detection. The peak current of Mb linearly decreased with the concentration of the complementary target DNA over a range from 0.3 to 300 nM with a detection limit of 0.1 nM. Furthermore, the selectivity of this biosensor for DNA hybridization was successfully examined.
引文
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