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>Fabrication of an Amperometric Flow-Injection Microfluidic Biosensor Based on Laccase for In Situ Determination of Phenolic Compounds
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Fabrication of an Amperometric Flow-Injection Microfluidic Biosensor Based on Laccase for In Situ Determination of Phenolic Compounds
We aim to develop an in situ microfluidic biosensor based on laccase from Trametes pubescens with flow-injection and amperometry as the transducer method. The enzyme was directly immobilized by potential step chronoamperometry, and the immobilization was studied using cyclic voltammetry and electrochemical impedance spectroscopy. The electrode response by amperometry was probed using ABTS and syringaldazine. A shift of interfacial electron transfer resistance and the electron transfer rate constant from 18.1 kΩ to 3.9 MΩ and 4.6 × 10−2 cm s−1 to 2.1 × 10−4 cm s−1, respectively, evidenced that laccase was immobilized on the electrode by the proposed method. We established the optimum operating conditions of temperature (55°C), pH (4.5), injection flow rate (200 µL min−1), and applied potential (0.4 V). Finally, the microfluidic biosensor showed better lower limit of detection (0.149 µM) and sensitivity (0.2341 nA µM−1) for ABTS than previous laccase-based biosensors and the in situ operation capacity.
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机译：我们的目标是开发一种基于Trametes pubescens漆酶的原位微流生物传感器，并采用流动注射和安培法作为传感器方法。通过电位步进计时安培法直接固定该酶，并使用循环伏安法和电化学阻抗谱研究固定化。使用ABTS和丁香醛嗪探测通过安培法测定的电极响应。界面电子转移电阻和电子转移速率常数从18.1kΩ到3.9MΩ和4.6×10 −2 cm s -1 转移到2.1×10 −4 cm s -1 分别证明了该方法将漆酶固定在电极上。我们确定了温度（55°C），pH（4.5），进样流速（200 µL min -1 ）和施加电势（0.4 V）的最佳操作条件。最后，与以前的基于漆酶的生物传感器相比，微流体生物传感器对ABTS的检测下限（0.149µm）和灵敏度（0.2341µnA µM -1 ）更好，并且具有更好的原位操作能力。
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