Due to the excessive contamination of heavy metals pollution, it is very urgent and necessary to develop a real-time detection method for the heavy metals in food. As a target sensing device, a paper-based microfluidic device (μPAD) has the advantages of simplicity, low-cost, and portability. In this study, a self-driven microfluidic paper-based chip was first developed for the simultaneous detection of four targets. The channels on the microfluidic chip were prepared by using wax printing and automatic screen printing on the filter paper, where liquid flowed by capillary force without pump assistance. Based on the specific binding ability of aptamers to heavy metals, a “turn-on” fluorescence aptasensor for the simultaneous detection of four heavy metal ions was developed on the proposed multi-channel device via smartphone imaging. The obtained fluorescence images were digitized into RGB color values by Image J software, and an M-mode was established to realize the quantitative detection of heavy metal ions. Under optimal conditions, the limits of detection for lead(II), mercury(II), cadmium(II), and arsenic(III) were 4.20 nM, 1.70 nM, 2.04 nM, and 1.65 nM, respectively. Furthermore, the aptasensor was successfully applied to the quantitative detection of four heavy metal ions in apple and lettuce samples with recovery rates of 84.0%–104.1%.
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