高效环烯烃共聚物芯片电泳的分析应用
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摘要
微芯片电泳是一种快速高效的微分析技术,有巨大的应用潜力,但由于芯片制作成本高、方法的实用性不足,目前在实际样品分析中的应用仍然相当有限。为了促进微芯片电泳的实际应用,本研究利用廉价的环烯烃共聚物(COC)塑料芯片,结合实验室自己搭建的低成本程控高压电源和激光诱导荧光检测装置,针对有重要现实意义的分析对象开展了以下几方面的工作：
第一章：介绍了微全分析系统的概念,简要总结了微芯片的发展历史、芯片制作材料、芯片制作技术、微芯片表面改性技术、微芯片分析的常用检测器以及微芯片电泳在实际样品分析中的应用。
第二章：建立了一种基于芯片电泳-激光诱导荧光检测的快速高效测定水样和服装样品中违禁芳香胺的方法。很多芳香胺有毒,甚至可致癌,故建立一种快速检测违禁芳香胺的方法很有必要。本实验利用含2%羟乙基纤维素(HPC)的10mmol/L的硼砂作为运行缓冲液,五种FITC标记的违禁芳香胺可以在90s内达到基线分离,检测限为1-3nmol/L,重复性好,理论塔板数可达6.8-8.5×105/m。该方法被成功用于水样和服装样品中违禁芳香胺的检测,回收率在85-110%之间。
第三章：建立了一种芯片电泳-激光诱导荧光间接检测苏丹红的方法。鉴于苏丹红染料的致癌性,许多国家已明令禁止在食品中添加苏丹红染料。苏丹红是偶氮染料,分解产物是芳香胺和萘酚类衍生物,本实验用两种不同的方法把苏丹红染料快速还原成芳香胺,然后用荧光胺标记,在激发波长为405nm的激光诱导荧光检测器上进行芯片电泳分离和检测。结果表明,四种苏丹红的还原产物可在60s内完成基线分离。
第四章：以HPC为添加剂对COC微芯片通道进行动态改性,建立了一种快速高效分离检测氧化型谷胱甘肽和还原型谷胱甘肽的方法。利用荧光胺为衍生试剂标记两种待测物,在405nm激光诱导荧光检测器上对样品进行检测,以含有1.0%HPC的10mmol/L的硼砂为运行缓冲液,两种谷胱甘肽可以在30s内完成基线分离,理论塔板数可达1.0×106/m。实验结果表明该方法简单、快速、重复性好,可用于人血清中谷胱甘肽的检测。
Microchip electrophoresis is a miniaturized analytical technique with rapid separation speed and high separation efficiency. But its application in real analysis is quite limited because of the high cost of chip fabrication, it has not been a practical method yet. In order to promote the practical application of microchip electrophoresis, cheap COC microchips combined with low-cost programmable high voltage power supplies and laser induced fluorescence detectors were used in our work for the analysis of some analytes with practical importance. There are four chapters in this thesis.
Chapter1:The μ-TAS was reviewed comprehensively. The development history of microfluidic chips, chip materials and production technologies, surface modification of microchannels, detection methods and the application of microchip-based electrophoresis in real samples were summarized.
Chapter2:A fast and efficient microchip electrophoretic method for the analysis of banned aromatic amines was established. Because some aromatic amines are strongly toxic and even carcinogenic, it is highly necessary to detect them rapidly. In this work,5banned aromatic amines that were labeled with fluorescein isothiocyanate could be baseline separated within90s by using10mmol/L borax containing2%(w/v) hydroxypropyl cellulose (HPC) as the running buffer. The limits of detection were in a range of1-3nmol/L and the theoretical plate numbers were in a range of6.8-8.5×105/m. The method was successfully applied in the analysis of these banned aromatic amines in waste water and textile samples, recoveries of added standards were85-110%.
Chapter3:A electrophoretic method using plastic microchips combined with low-cost laser-induced fluorescence detector was established to indirectly determinate sudan dyes, a group of dyes that are banned for use as food colorants due to their carcinogenicity. Because Sudan dyes can be reduced to aromatic amines and naphthalene phenolic derivatives, two methods of chemical reduction of Sudan dyes were adopted in this study. The aromatic amines from the dyes were labled by fluorescamine and were separated and detected with microchip electrophoresis with a405nm laser induced fluorescence detector. The experimental results showed that the reductive products could be baseline resolved within60s.
Chapter4:A microchip electrophoresis based method for fast and efficient separation and detection of oxidized and reduced glutathione was established. Using HPC as additive for dynamic modification of the microchannels of COC microchips, fluorescamine labeled oxidized and reduced glutathione could be baseline separated in30s. The number of theoretical plates is up to1.0×106/m. The method is simple, rapid and reproducible, and were successfully used for the detection of glutathione in human serum sample.
第一章：介绍了微全分析系统的概念,简要总结了微芯片的发展历史、芯片制作材料、芯片制作技术、微芯片表面改性技术、微芯片分析的常用检测器以及微芯片电泳在实际样品分析中的应用。
第二章：建立了一种基于芯片电泳-激光诱导荧光检测的快速高效测定水样和服装样品中违禁芳香胺的方法。很多芳香胺有毒,甚至可致癌,故建立一种快速检测违禁芳香胺的方法很有必要。本实验利用含2%羟乙基纤维素(HPC)的10mmol/L的硼砂作为运行缓冲液,五种FITC标记的违禁芳香胺可以在90s内达到基线分离,检测限为1-3nmol/L,重复性好,理论塔板数可达6.8-8.5×105/m。该方法被成功用于水样和服装样品中违禁芳香胺的检测,回收率在85-110%之间。
第三章：建立了一种芯片电泳-激光诱导荧光间接检测苏丹红的方法。鉴于苏丹红染料的致癌性,许多国家已明令禁止在食品中添加苏丹红染料。苏丹红是偶氮染料,分解产物是芳香胺和萘酚类衍生物,本实验用两种不同的方法把苏丹红染料快速还原成芳香胺,然后用荧光胺标记,在激发波长为405nm的激光诱导荧光检测器上进行芯片电泳分离和检测。结果表明,四种苏丹红的还原产物可在60s内完成基线分离。
第四章：以HPC为添加剂对COC微芯片通道进行动态改性,建立了一种快速高效分离检测氧化型谷胱甘肽和还原型谷胱甘肽的方法。利用荧光胺为衍生试剂标记两种待测物,在405nm激光诱导荧光检测器上对样品进行检测,以含有1.0%HPC的10mmol/L的硼砂为运行缓冲液,两种谷胱甘肽可以在30s内完成基线分离,理论塔板数可达1.0×106/m。实验结果表明该方法简单、快速、重复性好,可用于人血清中谷胱甘肽的检测。
Microchip electrophoresis is a miniaturized analytical technique with rapid separation speed and high separation efficiency. But its application in real analysis is quite limited because of the high cost of chip fabrication, it has not been a practical method yet. In order to promote the practical application of microchip electrophoresis, cheap COC microchips combined with low-cost programmable high voltage power supplies and laser induced fluorescence detectors were used in our work for the analysis of some analytes with practical importance. There are four chapters in this thesis.
Chapter1:The μ-TAS was reviewed comprehensively. The development history of microfluidic chips, chip materials and production technologies, surface modification of microchannels, detection methods and the application of microchip-based electrophoresis in real samples were summarized.
Chapter2:A fast and efficient microchip electrophoretic method for the analysis of banned aromatic amines was established. Because some aromatic amines are strongly toxic and even carcinogenic, it is highly necessary to detect them rapidly. In this work,5banned aromatic amines that were labeled with fluorescein isothiocyanate could be baseline separated within90s by using10mmol/L borax containing2%(w/v) hydroxypropyl cellulose (HPC) as the running buffer. The limits of detection were in a range of1-3nmol/L and the theoretical plate numbers were in a range of6.8-8.5×105/m. The method was successfully applied in the analysis of these banned aromatic amines in waste water and textile samples, recoveries of added standards were85-110%.
Chapter3:A electrophoretic method using plastic microchips combined with low-cost laser-induced fluorescence detector was established to indirectly determinate sudan dyes, a group of dyes that are banned for use as food colorants due to their carcinogenicity. Because Sudan dyes can be reduced to aromatic amines and naphthalene phenolic derivatives, two methods of chemical reduction of Sudan dyes were adopted in this study. The aromatic amines from the dyes were labled by fluorescamine and were separated and detected with microchip electrophoresis with a405nm laser induced fluorescence detector. The experimental results showed that the reductive products could be baseline resolved within60s.
Chapter4:A microchip electrophoresis based method for fast and efficient separation and detection of oxidized and reduced glutathione was established. Using HPC as additive for dynamic modification of the microchannels of COC microchips, fluorescamine labeled oxidized and reduced glutathione could be baseline separated in30s. The number of theoretical plates is up to1.0×106/m. The method is simple, rapid and reproducible, and were successfully used for the detection of glutathione in human serum sample.
引文
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