基于厚膜技术的血液生物化学传感器的研究
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摘要
近年来,高性能、低成本、微小型医疗仪器不断向传统医疗仪器发起挑战。一种新的临床检验模式——床边检测应运而生。本课题研究的手持式血液分析传感器是典型的床边检测设备,针对血液生化和电解质这些临床生化检验中最为基础和重要的内容,实现血液样本的即时测量。其快速、廉价和操作简单的特性,特别适合我国社区、农村诊所医疗现状和野外、急救的诊断需求,具有重要的社会价值和现实意义。
针对目前离子检测仪器体积大、操作复杂、成本昂贵的缺点,本课题研究了一种高性能、低成本的离子敏传感器制备技术。首次提出在丝网印刷碳糊电极基底上,采用厚膜成膜技术制备全固态离子选择性电极及其相应的外参比电极。通过大量实验,对厚膜成膜技术参数进行了优化,成功地实现了采用丝网印刷的方法制备固态导电聚合物PEDOT(PSS)层作为内充电解质、中性载体作为敏感物质、高分子材料聚氯乙烯为支持体的固态敏感膜的平板型全固态离子选择性电极。创新性的将参比电极设计为与工作电极类似的结构,利用干扰电位相减相消的原理提高传感器的抗干扰性能,并且缩短了传感器达到稳态的时间。同传统离子敏传感器相比,这种平板全固态离子敏传感器去除了液态部分,具有体积小易使用、携带保存方便的特点,特别适用于床边检测。对制备的钾、钙、钠、pH四种离子敏传感器进行了大量血清测量实验,考察了电极的灵敏度、一致性、保存期、环境影响等各方面性能,证明该四种离子敏传感器基本可以达到临床诊断的要求。
针对用于血液生化检测的葡萄糖传感器,研究了静电纺丝法和电聚合导电聚合物法固定葡萄糖氧化酶的技术,对各项制备参数进行了优化。另外进行了碳纳米管对传感器响应增强方面的研究,分别制备了采用新型PQQ-GDH酶作为敏感物质和采用静电纺丝聚丙烯腈/丙烯酸固定葡萄糖氧化酶的碳纳米管修饰的葡萄糖传感器。通过对比实验证明采用碳纳米管修饰的传感器,其灵敏度、电极反应速度都得到了极大改善。
In recent years, micro or small medical devices with high-performance and low-cost have been challenging conventional medical equipments. A new clinical testing mode-the point of care testing (POCT) came into being, because of its portability, easy operation and low cost. POCT is gradually becoming the trend of medical diagnosis. In this thesis, a typical POCT device-- a handheld bio-chemical sensor for blood electrolyte or glucose analysis was studied. It can perform a complete panel of tests, affording facilities for doctors to monitor and diagnose the respiratory status, blood acid-base balance, ion concentration and metabolism of their patients quickly and effectively. The portable blood analyzer has huge social needs and market prospects, particularly suitable for community and rural clinics, field and first aid.
Planar all-solid-state sensors based on thick film were developed to measure the concentration of K+, Ca2+, Na+ and pH in human serum. Each sensor consists of an ion-selective electrode and one external reference electrode. Experimentally, the parameters for thick film screen-printing were optimized. Successfully, polyvinyl chloride with neutral carries was formed as solid-state ion selective membrane, after conductive polymer PEDOT(PSS) was coated on the screen-printed carbon electrodes as inner solid-state electrolyte. A structure of external reference electrode similar to the working electrode was innovated to improve the anti-interference performance of the sensors based on the principle of interference potential cancellation. The design also speeded up the sensors to reach steady state. Compared with conventional ion selective electrodes, such planar all-solid-state sensors without liquid part, were easy to store and carry, especially suitable for bedside testing. Lots of clinic trials were carried out to test potassium, calcium, sodium, pH in serum using the sensors developed. The sensitivity, repeatability, reproducibility, stability, and environmental impact of the sensors were explored. The results indicated these four kinds of blood ion. sensors could meet the requirement of clinical diagnosis.
Enzyme immobilization is a crucial technology for biosensors using enzymes as sensitive materials. Electrospinning and electropolymerization methods were studied to immobilize glucose oxidase on Pt electrode and screen-printed carbon electrode, respectively. After the processing parameters were optimized, the resulted glucose sensors had low detection limit, good sensitivity and linear detection range, reached the clinical diagnosis requirement. In addition, the effect of carbon nanotubes on sensor response enhancement was studied. Based on PQQ-GDH, a carbon nanotubes modified glucose biosensor was developed. Based on glucose oxidase immobilized by electrospun PANCAA, another carbon nanotubes modified glucose biosensor was developed as well. Based on the experimental results showing that the sensitivity and the electrode reaction rate of the sensors modified by carbon nanotubes are significantly improved, the mechanism of the carbon nanotubes modification was analyzed.
针对目前离子检测仪器体积大、操作复杂、成本昂贵的缺点,本课题研究了一种高性能、低成本的离子敏传感器制备技术。首次提出在丝网印刷碳糊电极基底上,采用厚膜成膜技术制备全固态离子选择性电极及其相应的外参比电极。通过大量实验,对厚膜成膜技术参数进行了优化,成功地实现了采用丝网印刷的方法制备固态导电聚合物PEDOT(PSS)层作为内充电解质、中性载体作为敏感物质、高分子材料聚氯乙烯为支持体的固态敏感膜的平板型全固态离子选择性电极。创新性的将参比电极设计为与工作电极类似的结构,利用干扰电位相减相消的原理提高传感器的抗干扰性能,并且缩短了传感器达到稳态的时间。同传统离子敏传感器相比,这种平板全固态离子敏传感器去除了液态部分,具有体积小易使用、携带保存方便的特点,特别适用于床边检测。对制备的钾、钙、钠、pH四种离子敏传感器进行了大量血清测量实验,考察了电极的灵敏度、一致性、保存期、环境影响等各方面性能,证明该四种离子敏传感器基本可以达到临床诊断的要求。
针对用于血液生化检测的葡萄糖传感器,研究了静电纺丝法和电聚合导电聚合物法固定葡萄糖氧化酶的技术,对各项制备参数进行了优化。另外进行了碳纳米管对传感器响应增强方面的研究,分别制备了采用新型PQQ-GDH酶作为敏感物质和采用静电纺丝聚丙烯腈/丙烯酸固定葡萄糖氧化酶的碳纳米管修饰的葡萄糖传感器。通过对比实验证明采用碳纳米管修饰的传感器,其灵敏度、电极反应速度都得到了极大改善。
In recent years, micro or small medical devices with high-performance and low-cost have been challenging conventional medical equipments. A new clinical testing mode-the point of care testing (POCT) came into being, because of its portability, easy operation and low cost. POCT is gradually becoming the trend of medical diagnosis. In this thesis, a typical POCT device-- a handheld bio-chemical sensor for blood electrolyte or glucose analysis was studied. It can perform a complete panel of tests, affording facilities for doctors to monitor and diagnose the respiratory status, blood acid-base balance, ion concentration and metabolism of their patients quickly and effectively. The portable blood analyzer has huge social needs and market prospects, particularly suitable for community and rural clinics, field and first aid.
Planar all-solid-state sensors based on thick film were developed to measure the concentration of K+, Ca2+, Na+ and pH in human serum. Each sensor consists of an ion-selective electrode and one external reference electrode. Experimentally, the parameters for thick film screen-printing were optimized. Successfully, polyvinyl chloride with neutral carries was formed as solid-state ion selective membrane, after conductive polymer PEDOT(PSS) was coated on the screen-printed carbon electrodes as inner solid-state electrolyte. A structure of external reference electrode similar to the working electrode was innovated to improve the anti-interference performance of the sensors based on the principle of interference potential cancellation. The design also speeded up the sensors to reach steady state. Compared with conventional ion selective electrodes, such planar all-solid-state sensors without liquid part, were easy to store and carry, especially suitable for bedside testing. Lots of clinic trials were carried out to test potassium, calcium, sodium, pH in serum using the sensors developed. The sensitivity, repeatability, reproducibility, stability, and environmental impact of the sensors were explored. The results indicated these four kinds of blood ion. sensors could meet the requirement of clinical diagnosis.
Enzyme immobilization is a crucial technology for biosensors using enzymes as sensitive materials. Electrospinning and electropolymerization methods were studied to immobilize glucose oxidase on Pt electrode and screen-printed carbon electrode, respectively. After the processing parameters were optimized, the resulted glucose sensors had low detection limit, good sensitivity and linear detection range, reached the clinical diagnosis requirement. In addition, the effect of carbon nanotubes on sensor response enhancement was studied. Based on PQQ-GDH, a carbon nanotubes modified glucose biosensor was developed. Based on glucose oxidase immobilized by electrospun PANCAA, another carbon nanotubes modified glucose biosensor was developed as well. Based on the experimental results showing that the sensitivity and the electrode reaction rate of the sensors modified by carbon nanotubes are significantly improved, the mechanism of the carbon nanotubes modification was analyzed.
引文
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