CN112268943B - 微电极光电化学传感器及其制备方法与应用 - Google Patents
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Abstract
本发明属于光电化学技术领域，公开了一种微电极光电化学传感器及其制备方法与应用，微电极光电化学传感器包括金属微纳米电极，金属微纳米电极外层为玻璃毛细管，金属微纳米电极尖端端部电沉积有金属氧化物半导体薄层，金属氧化物半导体薄层尖端端部键合有有机光电分子层；其制备方法为将金属丝固定于毛细玻璃管内，经拉制、电沉积、热处理、硅烷偶联处理、键合等步骤，即得微电极光电化学传感器。本发明实现了对生物微环境中活性氧物种的原位实时高选择性检测，防止了生物分子的非特异性吸附，有效避免了其它活性物质的干扰。本发明的制备方法适用于制备微电极光电化学传感器，所制的微电极光电化学传感器适用于检测生物活性氧物种。
Description
技术领域
本发明属于光电化学技术领域，涉及一种传感器，具体地说是一种微电极光电化学传感器及其制备方法与应用。
背景技术
生物活性氧物种(ROS)，包括单线态氧、过氧化氢、羟基自由基、超氧自由基、氢过氧自由基、次氯酸等，在生物体内发挥着重要的作用，如参与许多生物信号的传导及炎症反应，但活性氧物种的过量生产会氧化破坏多种生物功能分子(如核酸、碳水化合物、脂质和蛋白质等)，导致分子和细胞功能丧失，此外，活性氧物种的含量还与多种疾病(如阿尔茨海默氏病(AD)、癌症、糖尿病、心脏病、心血管疾病等)密切相关。但是，在一些特殊的治疗方式中，如光动力治疗(PDT)、化学动力学疗法(CDP)等治疗方式，会利用活性氧物种的破坏作用来杀伤病变细胞及组织。由此可见，生物体中活性氧物种的检测具有重要意义，但是活性氧物种的寿命很短且扩散半径小，所以对它的时空分辨检测存在较大的挑战。
现有的活性氧物种检测方法主要有光谱法和基于微电极的电化学方法。光谱法具体包括电子顺磁共振法、瞬态光谱法、荧光成像法等，其中电子顺磁共振法与瞬态光谱法无法实现生物体的原位操作，而荧光成像法存在探针分子扩散而难以实现空间的准确定位。
微电极是指电极的一维尺寸为微米(10-6m)级至纳米(10-9m)级的一类电极，当电极的一维尺寸从毫米级降低至微米级时，表现出了许多优良的电化学特征，如极高的稳态电流密度、极短的响应时间、极化电流小、欧姆压降小、传质速度高、信噪比大等，可用于瞬态电极过程、高阻抗电解质体系和流动体系的研究。最近几十年以来，基于微电极的微型传感器逐渐开始成为新兴的电化学测量技术，微电极传感器具有良好的电化学性能和灵敏度高的特点，可以测定微纳米级的物质浓度水平。虽然基于微电极的电化学方法具有上述优势，但现有的微电极在复杂生物环境的检测中还存在诸多问题，如电极表面容易产生非特异性吸附，需要对电极施加偏压，但是施加偏压之后，其它活性物质容易在电极表面发生电子转移而产生较大干扰电流，难以实现不同类型活性氧物种的选择性检测。
发明内容
本发明的目的，是要提供一种微电极光电化学传感器，实现不同类型活性氧物种的选择性检测；本发明的另一个目的，是要提供上述微电极光电化学传感器的一种制备方法。
本发明为实现上述目的，所采用的技术方法如下：
一种微电极光电化学传感器的制备方法，按照以下步骤进行：
S1、将金属丝固定于毛细玻璃管内，拉制处理成金属微纳米电极；
S2、在金属微纳米电极尖端端部电沉积金属M，金属M经过氧化或热处理形成金属氧化物半导体薄层，之后采用3-氨丙基三乙氧基硅烷将金属氧化物半导体薄层表面进行硅烷偶联处理，即得微电极基底；
S3、微电极基底通过酰胺键与有机光电分子结合即得微电极光电化学传感器。
作为限定：所述金属丝材料为铂或金，金属丝直径为20～50微米，毛细玻璃管的材料为石英或硼硅酸盐，毛细玻璃管的内径为0.2～1毫米，金属微纳米电极的直径为0.2～10微米。
作为进一步限定：所述金属M为钛、铁、锡、镉、铬、铝中的一种，金属氧化物半导体薄层厚度为0.1～2微米。
作为再进一步限定：所述步骤S3中有机光电分子为
其中，R为
中的一种。
本发明还提供了上述制备方法制备的微电极光电化学传感器，它包括金属微纳米电极，金属微纳米电极外层为玻璃毛细管，金属微纳米电极尖端端部电沉积有金属氧化物半导体薄层，金属氧化物半导体薄层尖端端部键合有有机光电分子层。
本发明还提供了上述微电传感器的一种应用，所述微电极光电化学传感器用于检测生物活性氧物种。
作为限定：微电极光电化学传感器检测生物活性氧物种的具体检测方法为：1)将微电极光电化学传感器置于含生物活性氧物种的待测样品中，反应时间为1-60min；2)采用电化学工作站以三电极体系对待测样品进行测试，微电极光电化学传感器为工作电极，参比电极和对电极均采用铂丝电极；3)用时间-电流法对生物活性氧物种进行检测。
本发明由于采用了上述方案，与现有技术相比，所取得的有益效果是：
本发明将光电化学分析的手段引入微电极，组建了具有金属、半导体和有机光电分子多级结构的电极材料的微电极光电化学传感器，微电极具有小尺寸效应，实现了活性氧物种的原位实时检测，利用具有特定结构的有机光电分子对目标活性氧物种的高特异性响应，实现了对生物微环境中活性氧物种的高选择性检测，此外，微电极光电化学传感器的表面修饰有金属氧化物半导体材料，并进行了硅烷偶联处理，得到了氨基功能化的微电极基底，防止了生物分子的非特异性吸附，而且在光电化学检测过程以光作为激发源，无须对电极施加较高偏压，从而有效避免了其它活性物质的干扰。
本发明的制备方法适用于制备微电极光电化学传感器，所制的微电极光电化学传感器适用于检测生物活性氧物种。
附图说明
下面结合附图及具体实施例对本发明作更进一步详细说明。
图1为本发明实施例微电极光电化学传感器的制备工艺流程图；
图2为本发明实施例微电极光电化学传感器的结构示意图；
图3为本发明实施例A部分的放大结构示意图；
图4(a)为本发明实施例微电极光电化学传感器对单线态氧1O2的电流响应示意图；
图4(b)为本发明实施例微电极光电化学传感器对超氧阴离子自由基O2˙-的电流响应示意图；
图4(c)为本发明实施例微电极光电化学传感器对过氧化氢H2O2的电流响应示意图；
图4(d)为本发明实施例微电极光电化学传感器对氢氧自由基HO˙的电流响应示意图；
图中：1、毛细玻璃管；2、金属丝；3、金属微纳米电极；4、金属氧化物半导体薄层；5、有机光电分子层。
具体实施方式
下面结合实施例对本发明作进一步说明，但本领域的技术人员应当理解，本发明并不限于以下实施例，任何在本发明具体实施例基础上做出的改进和等效变化，都在本发明权利要求保护的范围之内。
实施例微电极光电化学传感器及其制备方法与应用
一种微电极光电化学传感器的制备方法，如图1所示，按照以下步骤进行：
S1、将金属丝2固定于毛细玻璃管1内，采用激光拉制仪将其拉制处理成金属微纳米电极3；
S2、在金属微纳米电极3尖端端部电沉积金属M，金属M经过氧化或热处理形成金属氧化物半导体薄层4，之后采用3-氨丙基三乙氧基硅烷(APTES)将金属氧化物半导体薄层4表面进行硅烷偶联处理，即得微电极基底；
S3、微电极基底通过化学键合作用与有机光电分子结合即得微电极光电化学传感器。
由上述制备方法制备的微电极光电化学传感器，如图2-图3所示，它包括金属微纳米电极3，金属微纳米电极3外层为玻璃毛细管1，金属微纳米电极3尖端端部电沉积有金属氧化物半导体薄层4，金属氧化物半导体薄层尖端端部键合有有机光电分子层5。
本实施例中金属丝材料为铂或金，金属M为钛、铁、锡、镉、铬、铝中的一种，有机光电分子为
其中，R为
中的一种。
微电极光电化学传感器用于检测生物活性氧物种，生物活性氧物种，包括单线态氧1O2、超氧阴离子自由基O2·-、过氧化氢H2O2、氢氧自由基HO·，具体的检测方法为：1)将微电极光电化学传感器置于含生物活性氧物种的待测样品中(溶液或细胞样品)，反应时间为1-60min；2)采用电化学工作站以三电极体系对待测样品进行测试，微电极光电化学传感器为工作电极，参比电极和对电极均采用铂丝电极；3)用时间-电流法对生物活性氧物种进行检测，电极偏压为-0.5-1.0V，利用微电极光电化学传感器使用样品处理前后的电流响应差值实现对样品活性氧物种的定量分析，微电极有机光电分子
中R为
检测的活性氧物种为单线态氧1O2，1O2能与蒽结构单元反应生成过氧化物，从而使得该结构单元的给电子能力减弱，光电流下降，如图4(a)所示，通过标准溶液测试建立光电流差值与1O2浓度的线性关系，进而对样品中1O2的定量分析；R为
检测的活性氧物种为超氧阴离子自由基O2·-，O2·-能与三氟甲磺酸酯基发生特异性解离反应，释放出酚羟基，具有较强供电子能力，从而电极光电流增强，如图4(b)所示，通过标准溶液测试建立光电流差值与O2·-浓度的线性关系，进而对样品中O2·-的定量分析；R为
检测的活性氧物种为过氧化氢H2O2，H2O2能进攻苯硼酸酯基团的硼原子中心形成不稳定的四取代中间产物，随后发生碳到硼酸盐中氧的迁移反应，最后水解形成具有较强给电子能力的酚羟基结构，从而电极光电流增强，如图4(c)所示，通过标准溶液测试建立光电流差值与H2O2浓度的线性关系，进而对样品中H2O2的定量分析；R为
检测的活性氧物种为氢氧自由基HO˙,HO˙能通过单电子氧化过程与苯氧基苯酚反应，从而使光敏分子游离出具有较强供电子能力的酚羟基，电极光电流增强，如图4(d)所示，通过标准溶液测试建立光电流差值与HO˙浓度的线性关系，进而对样品中HO˙的定量分析。
Claims (6)
1.一种微电极光电化学传感器的制备方法，其特征在于，按照以下步骤进行：
S1、将金属丝固定于毛细玻璃管内，拉制处理成金属微纳米电极；
S2、在金属微纳米电极尖端端部电沉积金属M，金属M经过氧化或热处理形成金属氧化物半导体薄层，之后采用3-氨丙基三乙氧基硅烷将金属氧化物半导体薄层表面进行硅烷偶联处理，即得微电极基底；
S3、微电极基底通过酰胺键与有机光电分子结合即得微电极光电化学传感器；其中，有机光电分子为，其中，R为、、、中的一种。
2.根据权利要求1所述的微电极光电化学传感器的制备方法，其特征在于，所述金属丝材料为铂或金，金属丝直径为20～50微米，毛细玻璃管的材料为石英或硼硅酸盐，毛细玻璃管的内径为0.2～1 毫米，金属微纳米电极的直径为0.2～10微米。
3.根据权利要求1或2所述的微电极光电化学传感器的制备方法，其特征在于，所述金属M为钛、铁、锡、镉、铬、铝中的一种，金属氧化物半导体薄层厚度为0.1～2微米。
4.权利要求1-3中任意一项所述制备方法制备的微电极光电化学传感器，其特征在于，它包括金属微纳米电极，金属微纳米电极外层为玻璃毛细管，金属微纳米电极尖端端部电沉积有金属氧化物半导体薄层，金属氧化物半导体薄层尖端端部键合有有机光电分子层。
5.权利要求4所述的微电极光电化学传感器的一种应用，其特征在于，所述微电极光电化学传感器用于检测生物活性氧物种。
6.根据权利要求5所述的应用，其特征在于，微电极光电化学传感器检测生物活性氧物种的具体检测方法为：1）将微电极光电化学传感器置于含生物活性氧物种的待测样品中，反应时间为1-60 min；2）采用电化学工作站以三电极体系对待测样品进行测试，微电极光电化学传感器为工作电极，参比电极和对电极均采用铂丝电极；3）用时间-电流法对生物活性氧物种进行检测。
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