Fluorescent aptasensor mediated with multiple ssDNA for sensitive detection of acetamiprid in vegetables based on magnetic Fe3O4/C-assisted separation

Abstract

Acetamiprid (ACE) is a highly effective broad-spectrum insecticide, and its widespread use is potentially harmful to human health and environmental safety. In this study, magnetic Fe₃O₄/carbon (Fe₃O₄/C), a derivative of metal-organic framework MIL-101 (Fe), was synthesized by a two-step calcination method. And a fluorescent sensing strategy was developed for the efficient and sensitive detection of ACE using Fe₃O₄/C and multiple complementary single-stranded DNA (ssDNA). By using aptamer with multiple complementary ssDNA, the immunity of interference of the aptasensor was improved, and the aptasensor showed high selectivity and sensitivity. When ACE was present, the aptamer (Apt) combined with ACE. The complementary strand of Apt (Cs1) combined with two short complementary strands of Cs1, fluorophore 6-carboxyfluorescein-labeled complementary strand (Cs2-FAM) and the other strand Cs3. The three strands formed a double-stranded structure, and fluorescence would not be quenched by Fe₃O₄/C. In the absence of ACE, Cs2-FAM would be in a single-chain state and would be adsorbed by Fe₃O₄/C, and the fluorescence of FAM would be quenched by Fe₃O₄/C via photoelectron transfer. This aptasensor sensitively detected ACE over a linear concentration range of 10-1000 nM with a limit of detection of 3.41 nM. The recoveries of ACE spiked in cabbage and celery samples ranged from 89.49% to 110.76% with high accuracy.

Keywords: Acetamiprid; Aptasensor; Metal–organic framework; Photoelectron transfer.