组成图示
示意图生成中
传感器类型
荧光生物传感器
检测对象
阿特拉津（atrazine）、异丙隆（isoproturon）、雌酮（estrone）；样品基质为饮用水厂各处理阶段水样（河水、砂滤水、臭氧水、活性炭滤水、饮用水）
检测原理
该传感器采用间接竞争免疫分析。样品中的阿特拉津、异丙隆或雌酮与Cy5.5标记的相应抗体孵育，形成抗原-抗体复合物；未结合的荧光抗体随后结合到玻璃芯片上空间分辨固定的氨基葡聚糖-分析物衍生物捕获区。分析物浓度越高，能结合到芯片表面的荧光抗体越少，表面荧光强度越低。He-Ne激光在玻璃芯片中发生全内反射，产生倏逝场并激发表面Cy5.5荧光；荧光经光纤收集、滤光后由光电二极管锁相检测。三个独立捕获区可同步输出三种污染物的信号，无需酶催化或核酸放大，信号直接来自荧光标记抗体。
检测灵敏度
LOD (µg/l): estrone 0.080; atrazine 0.160; isoproturon 0.050；线性范围 (µg/l): estrone 0.170–10.7; atrazine 0.350–1.47; isoproturon 0.110–2.83；R^2: estrone 0.992; atrazine 0.979; isoproturon 0.998
效应效果
与SPE-LC-MS相比，RIANA在线性、灵敏度和准确度上较低：LC-MS回收率为94–100%，RIANA为117–175%；但RIANA重复性更好，RSD为1–7%，LC-MS为5–15%。交叉反应导致高估，如异丙隆测定中diuron、linuron、chlortoluron交叉反应分别为92.8%、53.4%、46.3%，阿特拉津测定中desethylatrazine、simazine、deisopropylatrazine为42.8%、19%、18.7%。加入OVA可减轻基质干扰，换能器可完成至少300次测量，单周期15 min，每天可分析约25个样品。作者认为其适合水厂污染物的快速、低成本、自动化筛查。
传感器的构成
- 基底/换能器：玻璃干涉层（glass interference layer，58 mm×10 mm×1.5 mm，45°斜面），作为TIRF光波导与光学换能芯片
- 表面活化层：3-缩水甘油氧基丙基三甲氧基硅烷（GOPTs）硅烷化，提供环氧基活化表面
- 识别捕获层：氨基葡聚糖-分析物衍生物偶联物（aminodextran-atrazine、aminodextran-estrone、aminodextran-isoproturon），空间分辨共价固定于三个独立区域
- 信号标记物：抗雌酮、抗异丙隆、抗阿特拉津抗体（anti-estrone、anti-isoproturon、anti-atrazine antibodies），用Cy5.5荧光染料标记
- 封闭剂：卵白蛋白（OVA），加入样品以饱和非特异结合位点、降低基质干扰
- 流动与再生系统：流动注射系统（FIA）、六通阀、1 mL注射泵、流动池；SDS再生液（0.5 g/100 mL，pH 1.9）清洗再生
- 光学读出系统：He-Ne激光（633 nm，7 mW）、光纤、滤光片、光电二极管和锁相检测，采集TIRF荧光信号
中文摘要
本文报道了光学多分析物免疫传感器RIANA在真实水样中同时检测三种环境有机污染物的应用，包括农药阿特拉津（atrazine）、异丙隆（isoproturon）和雌激素雌酮（estrone）。该传感器基于化学修饰光学换能芯片上的间接抑制免疫分析，通过空间分辨表面修饰和全内反射荧光（TIRF）实现多目标同步测定。作者将免疫传感器方法与固相萃取-液相色谱-质谱（SPE-LC-MS）进行验证比较。结果表明，色谱法在线性、灵敏度和准确度方面更优，而生物传感器在重复性、速度、成本和自动化方面具有优势。两种方法平行用于监测水厂砂滤、臭氧、活性炭和氯化处理过程中污染物的存在与去除。生物传感器结果存在一定高估，部分归因于基质效应和交叉反应，尽管加入卵白蛋白可减少基质干扰。总体而言，RIANA可作为无需样品前处理的快速、简便、自动化水污染物筛查工具，这也是其多分析物配置用于水厂常规监测的首次报道。
英文摘要
This work describes the application of an optical biosensor (RIver ANALyser, RIANA) to the simultaneous analysis of three relevant environmental organic pollutants, namely, the pesticides atrazine and isoproturon and the estrogen estrone, in real water samples. This biosensor is based on an indirect inhibition immunoassay which takes place at a chemically modified optical transducer chip. The spatially resolved modification of the transducer surface allows the simultaneous determination of selected target analytes by means of "total internal reflection fluorescence" (TIRF). The performance of the immunosensor method developed was evaluated against a well accepted traditional method based on solid-phase extraction followed by liquid chromatography-mass spectrometry (LC-MS). The chromatographic method was superior in terms of linearity, sensitivity and accuracy, and the biosensor method in terms of repeatability, speed, cost and automation. The application of both methods in parallel to determine the occurrence and removal of atrazine, isoproturon and estrone throughout the treatment process (sand filtration, ozonation, activated carbon filtration and chlorination) in a waterworks showed an overestimation of results in the case of the biosensor, which was partially attributed to matrix and cross-reactivity effects, in spite of the addition of ovalbumin to the sample to minimize matrix interferences. Based on the comparative performance of both techniques, the biosensor emerges as a suitable tool for fast, simple and automated screening of water pollutants without sample pretreatment. To the author's knowledge, this is the first description of the application of the biosensor RIANA in the multi-analyte configuration to the regular monitoring of pollutants in a waterworks.