Automatable Flow System for Paraoxon Detection with an Embedded Screen-Printed Electrode Tailored with Butyrylcholinesterase and Prussian Blue Nanoparticles

Chemosensors

Fabiana Arduini¹  Daniela Neagu¹  Viviana Scognamiglio²  Sabrina Patarino¹  Danila Moscone¹  Giuseppe Palleschi¹  Paolo Ugo³ 

[1] Dipartimento di Scienze e Tecnologie Chimiche, Università di Roma Tor Vergata, Via della Ricerca Scientifica, Rome 00133, Italy; E-Mails:;Dipartimento di Scienze Bio-Agroalimentari, Istituto di Cristallografia (IC-CNR), Via Salaria km 29.300, 00015 Monterotondo, Rome 00015, Italy; E-Mail:Dipartimento di Scienze e Tecnologie Chimiche, Università di Roma Tor Vergata, Via della Ricerca Scientifica, Rome 00133, Italy;

关键词: screen-printed electrode;    biosensor;    inhibition;    butyrylcholinesterase;    paraoxon;    flow analysis;   

DOI  :  10.3390/chemosensors3020129

来源: mdpi

【 摘 要 】

Nowadays extensive volumes of pesticides are employed for agricultural and environmental practices, but they have negative effects on human health. The levels of pesticides are necessarily restricted by international regulatory agencies, thus rapid, cost-effective and in-field analysis of pesticides is an important issue. In the present work, we propose a butyrylcholinesterase (BChE)-based biosensor embedded in a flow system for organophosphorus pesticide detection. The BChE was immobilized by cross-linking on a screen-printed electrode modified with Prussian Blue Nanoparticles. The detection of paraoxon (an organophosphorus pesticide) was carried out evaluating its inhibitory effect on BChE, and quantifying the enzymatic hydrolysis of butyrylthiocholine before and after the exposure of the biosensor to paraoxon, by measuring the thiocholine product at a working voltage of +200 mV. The operating conditions of the flow system were optimized. A flow rate of 0.25 mL/min was exploited for inhibition steps, while a 0.12 mL/min flow rate was used for substrate measurement. A substrate concentration of 5 mM and an incubation time of 10 min allowed a detection limit of 1 ppb of paraoxon (corresponding to 10% inhibition). The stability of the probe in working conditions was investigated for at least eight measurements, and the storage stability was evaluated up to 60 days at room temperature in dry condition. The analytical system was then challenged in drinking, river and lake water samples. Matrix effect was minimized by using a dilution step (1:4 v/v) in flow analysis. This biosensor, embedded in a flow system, showed the possibility to detect paraoxon at ppb level using an automatable and cost-effective bioanalytical system.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.