【作者】 王军涛；

【导师】 黄俊；

【作者基本信息】 武汉理工大学 ， 光电子及信息材料学， 2006， 硕士

【摘要】 光纤传感器技术是当今世界最重要的检测技术之一,光纤传感器具有灵敏度高、响应快、操作简便、易于微型化等优点。光纤生物传感器是光纤传感器中的一个重要分支,在医学、生物工程、食品工业和环境监控等方面有广阔的应用前景。肾上腺素是一种非常重要的生理学神经递质,与人们的健康密切相关,人体内肾上腺素的检测,对常见疾病的诊断与治疗有重要意义,因此光纤肾上腺素传感器在临床诊断,生物技术方面有着广泛的应用前景。目前对于肾上腺素的各种检测方法都存在灵敏度不高,操作复杂等缺点,因此非常需要开发新型的光纤肾上腺素传感器。 光学生物敏感材料是光纤生物传感器中一个非常重要的部分,其性能直接影响光纤生物传感器的性能。有机-无机纳米复合材料由于其结构特殊、性能优异而成为材料学中的研究热点。有机—无机纳米复合粒子兼顾有机物和无机纳米粒子的诸多优点,且稳定性较好,因此被大量应用于生物物质的固定。 本文制备了一种优良的载体材料四氨基酞菁钴（CoTAPc）—Fe₃O₄纳米复合粒子,并以之固定了漆酶,所得到的固定化漆酶具有优良的特性。设计和构建了基于漆酶催化的肾上腺素传感器,对肾上腺素进行了初步的检测。 本论文的主要工作包括以下几个方面: （1）利用原位复合法制备了CoTAPc—Fe₃O₄纳米复合粒子。对所得的粒子利用红外、振动样品磁强计、扫描电镜等进行表征。所得到的纳米复合粒子的平均粒径可达到70nm左右,接近超顺磁性,分散性好,有良好的生物相容性,而且表面有大量活性官能团（-NH₂）,是一种优良的载体材料。 （2）以CoTAPc—Fe₃O₄纳米复合粒子为载体通过交联法固定了血红密孔菌真菌漆酶,研究了戊二醛的浓度、牛血清白蛋白的浓度、反应过程中的pH值、漆酶的用量等因素对固定化漆酶活性的影响,以ABTS为底物固定化漆酶的最佳使用温度为45℃,最佳使用pH值为3,Km值为18.3μM,比游离酶略大（12.6μM）。固定化漆酶具有较好的热稳定性、贮存稳定性和操作稳定性。 （3）研究了漆酶在不同条件下催化肾上腺素氧化的反应情况,在酸性条件下,漆酶催化肾上腺素氧化反应生成物为肾上腺素红,而在碱性条件下反应较为复杂,在pH等于9时,生成物主要为肾上腺素黄,且反应较快,这为在碱性条件下检测肾上腺素打下了基础。 （4）设计和构建了基于荧光猝灭原理的光纤肾上腺素传感检测系统,对肾上腺素进行了初步的检测,研究了其传感特性。更多还原

【Abstract】 Fiber optic sensor is one of the most important detection instruments. Fiber optic sensor has many advantages such as high sensitivity, quick response, easy operation and micromation. Fiber optic biosensor is an important branch of fiber optic sensor, and it has a wide application prospect in medical science, biological engineering, food industries and environment monitoring. Epinephrine is a crucial physiological neurotransmitter and is closely related to human health. As for the determination of epinephrine in the field of clinical diagnosis and biologic technology, a fiber optic biosensor is considered to be very necessary. The existing traditional methods for the determination of epinephrine have such deficiencies as low sensitivity or complicated operation that can’t meet the need of practical use. Therefore, it’s necessary to develop a novel fiber optic epinephrine biosensor.Biosensitive material is an important part of fiber optic biosensor. The characters of the material influence the whole fiber optic biosensor greatly. Organic-inorganic nanaparticle composite is a novel interdiscipline, it has good application in the future. Organic-inorganic nanaparticle composite has both the advantage of organic substance and inorganic nanocomposite. It has a wide application in immobilization of biological substances.In this thesis, a new kind of carrier cobalt tetraaminophthalocyanine （CoTAPc）-Fe₃O₄ with excellent characters was successfully prepared and was used in laccase immobilization. The immobilized laccase has good excellent performance. The fiber optic epinephrine sensor was designed and constructed, and several tests were performed to detect the concentration of epinephrine in solution.The work of this paper includes mainly four aspects as followed:（1） The CoTAPc-Fe₃O₄ nanoparticle composite was prepared by in situ complex technology . The composite were characterized by IR, FEG-SEM, XRD, EDS and Model 4HF VSM. The nanoparticle composites took the shape of roundish spheres with their mean diameter of about 70 nm. The nanoparticle composites have good dispersion, excellent compatibility and abundant active functions on the surface for immobilization of laccase.（2） The laccase was immobilized on the surface of the CoTAPc-Fe₃O₄ nanoparticle composite by crosslinking method. The effects of glutaraldehyde concentration, BSA concentration, the pH on the reaction process and the amount of laccase on the activity of immobilized laccase were studied. After immobilization, the best pH, temperature and Km of immobilized laccase catalyse ABTS were studied . The immobilized laccase has better stabilities in heat, storage and operation.（3） The reactive complexion of epinephrine’s oxygenation in different pH were studied. When pH<7, the product was adrenochrome. When pH>7, the reaction was complicated. The product was adrenolutine at pH=9. This work provided a basis of epinephrine’s detection in alkalescence.（4） A fiber optic epinephrine biosensor based on fluorescence quenching was designed and fabricated. The performance of this sensor in detecting epinephrine was investigated.更多还原