Electrochemical biotoxicity detection on a microfluidic paper-based analytical device via cellular respiratory inhibition

A novel microfluidic paper-based analytical device (μPAD) was developed with benzoquinone (BQ)-mediated E. coli respiration method to measure the biotoxicities of pollutants. Functional units including sample injection, fluid-cell separation, all-carbon electrode-enabled electrochemical detection, were integrated on a piece of chromatography paper. The three-electrode, working electrode, counter electrode and reference electrode, were simultaneously screen-printed on the μPAD with conductive carbon ink. The satisfying electrochemical performance of the paper-based carbon three-electrode was confirmed by cyclic voltammetry detecting K3 [Fe(CN)6]. The process of cell toxication was considered that toxicants inhibited cell respiration and diminished the electrons on E. coli respiratory chain. It was quantitatively reflected by measuring oxidation current of hydroquinone (HQ) as a reduced state of the redox mediator BQ after the incubation of cells with pollutants. The current detection time, BQ concentration and E. coli incubation time were carefully optimized to establish the systematic optimized operations of BQ-mediated E. coli respiration method. Using the fabricated μPAD the half inhibitory concentration (IC50) were Cu2+ solution 13.5 μg mL-1, Cu2+-soil 21.4 mg kg-1, penicillin sodium-soil 85.1 mg kg-1, and IC30 of Pb2+ solution was 60.0 μg mL-1. Detection of pesticide residues in vegetable juices were accomplished in a similar way. The proposed method is fascinating on three points; 1) The generality in the biotoxicity detection depends on toxicants inducing cellular respiratory inhibition; 2) The portability and affordability make it convenient for practical applications, because of replacing incubators and centrifuges; 3) There is potential applicability in less-developed areas due to its simple operation and low-cost.