Abstract In this work we report a novel paper-based analytical device read-out via LED-induced fluorescence detection (FPAD) for the quantification of the emerging pollutant ethinylestradiol (EE2) in river water samples. The PAD was used as a reaction platform for a competitive enzyme immunoassay. For the PAD development, microzones of filter paper, printed by a wax printing method, were modified with amino-functionalized SBA-15 and subsequently, anti-EE2 specific antibodies were covalently immobilized. The determination of EE2 in water was carried out by adding a fixed concentration of EE2 conjugated with the enzyme horseradish peroxidase (HRP) to samples and standards. Then, the FPAD were added and incubated for 10 min. Finally, the detection was performed by the reaction of 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) whose oxidation is catalyzed by HRP in the presence of H2O2, obtaining the highly fluorescent resorufin (R). Resorufin was detected by LED excitation at 550 nm, observing emission at 585 nm. The EE2 concentration in the samples was inversely proportional to the relative fluorescence obtained from the enzymatic reaction products. The FPAD assay showed a detection limit (LOD) of 0.05 ng L−1 and coefficients of variation (CV) below 4.5% within-assay and below 6.5% between-assay, respectively. The results obtained show the potential suitability of our FPAD for the selective and sensitive quantification of EE2 in river water samples. In addition, it has the PADs advantages of being disposable, easy to apply and inexpensive.

中文翻译：

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使用基于荧光纸的免疫传感器定量饮用水源中的炔雌醇

摘要 在这项工作中，我们报告了一种通过 LED 诱导荧光​​检测 (FPAD) 读出的新型纸质分析设备，用于量化河水样品中新兴污染物乙炔雌二醇 (EE2)。PAD 用作竞争性酶免疫分析的反应平台。对于 PAD 开发，通过蜡印法印刷的滤纸微区用氨基功能化的 SBA-15 进行修饰，随后共价固定抗 EE2 特异性抗体。通过向样品和标准品中加入固定浓度的与辣根过氧化物酶 (HRP) 偶联的 EE2 来测定水中的 EE2。然后，加入FPAD并孵育10分钟。最后通过10-乙酰-3的反应进行检测，7-二羟基吩恶嗪 (ADHP) 在 H2O2 存在下由 HRP 催化氧化，获得高荧光试卤灵 (R)。试卤灵在 550 nm 处通过 LED 激发检测，在 585 nm 处观察发射。样品中的 EE2 浓度与从酶促反应产物中获得的相对荧光成反比。FPAD 检测显示检测限 (LOD) 为 0.05 ng L-1，变异系数 (CV) 分别低于 4.5% 的检测内和低于 6.5% 的检测间。获得的结果表明我们的 FPAD 可能适用于河水样品中 EE2 的选择性和灵敏定量。此外，它具有一次性、易于应用和价格低廉的 PAD 优点。试卤灵在 550 nm 处通过 LED 激发检测，在 585 nm 处观察发射。样品中的 EE2 浓度与从酶促反应产物中获得的相对荧光成反比。FPAD 检测显示检测限 (LOD) 为 0.05 ng L-1，变异系数 (CV) 分别低于 4.5% 的检测内和低于 6.5% 的检测间。获得的结果表明我们的 FPAD 可能适用于河水样品中 EE2 的选择性和灵敏定量。此外，它具有一次性、易于应用和价格低廉的 PAD 优点。试卤灵在 550 nm 处通过 LED 激发检测，在 585 nm 处观察发射。样品中的 EE2 浓度与从酶促反应产物中获得的相对荧光成反比。FPAD 检测显示检测限 (LOD) 为 0.05 ng L-1，变异系数 (CV) 分别低于 4.5% 的检测内和低于 6.5% 的检测间。获得的结果表明我们的 FPAD 可能适用于河水样品中 EE2 的选择性和灵敏定量。此外，它具有一次性、易于应用和价格低廉的 PAD 优点。FPAD 检测显示检测限 (LOD) 为 0.05 ng L-1，变异系数 (CV) 分别低于 4.5% 的检测内和低于 6.5% 的检测间。获得的结果表明我们的 FPAD 可能适用于河水样品中 EE2 的选择性和灵敏定量。此外，它具有一次性、易于应用和价格低廉的 PAD 优点。FPAD 检测显示检测限 (LOD) 为 0.05 ng L-1，变异系数 (CV) 分别低于 4.5% 的检测内和低于 6.5% 的检测间。获得的结果表明我们的 FPAD 可能适用于河水样品中 EE2 的选择性和灵敏定量。此外，它具有一次性、易于应用和价格低廉的 PAD 优点。