荧光型光学复合生物敏感材料的制备与性能研究
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摘要
人体血液中胆固醇和葡萄糖浓度的检测对人体健康监测和疾病早期诊断十分重要,基于酶催化的光纤生物传感器是实现这类检测的有效手段之一。研制高性能的敏感材料是获得高性能光纤生物传感器的关键技术。目前报道的基于酶催化的光纤生物传感器的敏感材料通常由光敏感材料和生物酶两部分组成,如果将光学指示剂和酶通过合适的技术组合在一起制成光学复合敏感材料,可有效提高传感器的响应速度和检测精度,并可实现传感器的微型化,具有较高的应用价值。
本论文的工作主要包括以下两个主要方面:溶胶凝胶光学胆固醇复合敏感材料的制备及性能研究;化学交联法制备光学葡萄糖复合敏感材料的初步研究。主要内容如下:
(1)研究了溶胶—凝胶过程前驱体参数配比、干燥速率、基片、甲酰胺用量等制备因素对光学胆固醇复合敏感材料性能的影响,还研究了给酶量对光学胆固醇复合敏感材料酶活性的影响,并对光学胆固醇复合敏感材料进行了FTIR分析。
(2)研究了溶胶凝胶光学胆固醇复合敏感材料的催化曲线、最适温度、最适使用pH值、固定化酶活稳定性及指示剂包埋稳定性。该光学胆固醇复合敏感材料最适使用pH值为7.0,最适催化温度为40℃;其固定化酶活稳定性及指示剂包埋稳定性比较好。设计和构建了光纤胆固醇传感器,该传感器对胆固醇溶液浓度具有较好的线性关系,线性检测范围为100—500 mg/dl,响应时间为40 s左右,长期稳定性比较好。
(3)选用醋酸纤维素作为载体,钌联吡啶作为荧光指示剂,对利用化学交联法制备同时固定有葡萄糖氧化酶和荧光指示剂的光学葡萄糖复合敏感材料进行了初步的研究。初步探讨了活化条件、戊二醛、葡萄糖氧化酶的浓度、固定化时间等制备条件对光学葡萄糖复合敏感材料酶活性的影响。
(4)初步探讨了反应条件对固定化葡萄糖氧化酶活性的影响,光学葡萄糖复合敏感材料最适催化温度为38℃;最适使用pH值为7.0。设计和构建了光纤葡萄糖传感器测试系统,初步研究表明:传感器的检测范围为100—600 mg/dl,响应时间为20 s左右。
The detection of cholesterol and glucose concentration in blood is of great significance for monitoring human health and early diagnosis of diseases.Among the various detecting methods,fiber optic biosensor based on enzyme catalysis is an effective means.The research on high-performance optical sensing materials is the key technology to develop the fiber optic biosensors with good properties.The reported sensing materials for fiber optic biosensor based on enzyme catalysis usually consist of optical sensing part and catalysis part.If these two parts are combined into one using an appropriate method to form the opitcal complex sensing materials,the response rate and detecting precision of the sensor can be improved greatly,and the sensor micromation will be realized.This kind of work is very important to the application of fiber optic sensor based on enzyme catalysis.
The research work of this thesis mainly consist of two parts:preparation of cholesterol complex optic sensing materials by sol-gel method and research on its properties,preliminary study on preparing glucose complex sensing materials by chemical cross-linking method.Following is the main content:
(1) The influences of preparation conditions on the properties of the cholesterol sensitive materials have been studied,such as precursor ratio in the sol-gel process,drying rate,substrate and amount of formamide.The relationship between the enzyme activity of sensitive materials and the amount of enzyme has also been studied.FTIR analysis is used for the characterization of the materials.
(2) The properties of the optical complex cholesterol sensing materials,such as catalytic curve,optimal temperature,optimal pH value,stability of immobilized enzyme and and embedded indicator,have been studied.The optimal pH value is 7.0,optimal temperature is 40℃.The stability of immobilized enzyme and embedded indicator is good.Fiber optic cholesterol sensor is designed and built. This sensor shows a good linear relationship between the phase delay and the cholesterol concentration in the range of 100-500 mg/dl.It has good long-term stability and the response time of 40s.
(3) Initial study on the optical complex glucose sensiting materials containing both glucose oxidase and fluorescence indicator has been performed,using cellulose acetate as the carrier and ruthenium bipyridyl as the indicator.The cross-linking method was used to immobilize the enzyme.The factors such as activation condition,amount of glutaraldehyde,concentration of GOD,immobilized time to the enzyme activity of the sensing materials have been investigated.
(4) Preliminary study on the influences of reaction conditions to the activities of immobilized glucose oxidase have been carried out.The optimal catalytic temperature and pH value are 38℃and 7.0,respectively.The fiber optic glucose sensor with the optical complex glucose sensiting materials was studied.The results show that its detecting range is 100-600 mg/dl and its response time is 20s.
本论文的工作主要包括以下两个主要方面:溶胶凝胶光学胆固醇复合敏感材料的制备及性能研究;化学交联法制备光学葡萄糖复合敏感材料的初步研究。主要内容如下:
(1)研究了溶胶—凝胶过程前驱体参数配比、干燥速率、基片、甲酰胺用量等制备因素对光学胆固醇复合敏感材料性能的影响,还研究了给酶量对光学胆固醇复合敏感材料酶活性的影响,并对光学胆固醇复合敏感材料进行了FTIR分析。
(2)研究了溶胶凝胶光学胆固醇复合敏感材料的催化曲线、最适温度、最适使用pH值、固定化酶活稳定性及指示剂包埋稳定性。该光学胆固醇复合敏感材料最适使用pH值为7.0,最适催化温度为40℃;其固定化酶活稳定性及指示剂包埋稳定性比较好。设计和构建了光纤胆固醇传感器,该传感器对胆固醇溶液浓度具有较好的线性关系,线性检测范围为100—500 mg/dl,响应时间为40 s左右,长期稳定性比较好。
(3)选用醋酸纤维素作为载体,钌联吡啶作为荧光指示剂,对利用化学交联法制备同时固定有葡萄糖氧化酶和荧光指示剂的光学葡萄糖复合敏感材料进行了初步的研究。初步探讨了活化条件、戊二醛、葡萄糖氧化酶的浓度、固定化时间等制备条件对光学葡萄糖复合敏感材料酶活性的影响。
(4)初步探讨了反应条件对固定化葡萄糖氧化酶活性的影响,光学葡萄糖复合敏感材料最适催化温度为38℃;最适使用pH值为7.0。设计和构建了光纤葡萄糖传感器测试系统,初步研究表明:传感器的检测范围为100—600 mg/dl,响应时间为20 s左右。
The detection of cholesterol and glucose concentration in blood is of great significance for monitoring human health and early diagnosis of diseases.Among the various detecting methods,fiber optic biosensor based on enzyme catalysis is an effective means.The research on high-performance optical sensing materials is the key technology to develop the fiber optic biosensors with good properties.The reported sensing materials for fiber optic biosensor based on enzyme catalysis usually consist of optical sensing part and catalysis part.If these two parts are combined into one using an appropriate method to form the opitcal complex sensing materials,the response rate and detecting precision of the sensor can be improved greatly,and the sensor micromation will be realized.This kind of work is very important to the application of fiber optic sensor based on enzyme catalysis.
The research work of this thesis mainly consist of two parts:preparation of cholesterol complex optic sensing materials by sol-gel method and research on its properties,preliminary study on preparing glucose complex sensing materials by chemical cross-linking method.Following is the main content:
(1) The influences of preparation conditions on the properties of the cholesterol sensitive materials have been studied,such as precursor ratio in the sol-gel process,drying rate,substrate and amount of formamide.The relationship between the enzyme activity of sensitive materials and the amount of enzyme has also been studied.FTIR analysis is used for the characterization of the materials.
(2) The properties of the optical complex cholesterol sensing materials,such as catalytic curve,optimal temperature,optimal pH value,stability of immobilized enzyme and and embedded indicator,have been studied.The optimal pH value is 7.0,optimal temperature is 40℃.The stability of immobilized enzyme and embedded indicator is good.Fiber optic cholesterol sensor is designed and built. This sensor shows a good linear relationship between the phase delay and the cholesterol concentration in the range of 100-500 mg/dl.It has good long-term stability and the response time of 40s.
(3) Initial study on the optical complex glucose sensiting materials containing both glucose oxidase and fluorescence indicator has been performed,using cellulose acetate as the carrier and ruthenium bipyridyl as the indicator.The cross-linking method was used to immobilize the enzyme.The factors such as activation condition,amount of glutaraldehyde,concentration of GOD,immobilized time to the enzyme activity of the sensing materials have been investigated.
(4) Preliminary study on the influences of reaction conditions to the activities of immobilized glucose oxidase have been carried out.The optimal catalytic temperature and pH value are 38℃and 7.0,respectively.The fiber optic glucose sensor with the optical complex glucose sensiting materials was studied.The results show that its detecting range is 100-600 mg/dl and its response time is 20s.
引文
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