Biomimetic Enzyme-linked Immunosorbent Assay Using a Hydrophilic Molecularly Imprinted Membrane for Recognition and Fast
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Biomimetic Enzyme-linked Immunosorbent Assay Using a Hydrophilic Molecularly Imprinted Membrane for Recognition and Fast Determination of Trichlorfon and Acephate Residues in Vegetables Chen Shi & Xueyan Liu & Lingyu Song & Xuguang Qiao & Zhixiang Xu
Received: 10 December 2014 / Accepted: 27 February 2015 # Springer Science+Business Media New York 2015
Abstract This study describes the development of a new direct competitive enzyme-linked immunosorbent assay method with a hydrophilic imprinted film as biomimetic antibody for determining multi-pesticide residues. The imprinted film, which can selectively recognize trichlorfon and acephate, was synthesized directly on the surface of a 96-well plate using 4-(dimethoxyphosphorothioylamino) butanoic acid as the template molecule. This film exhibited antibody-like recognition ability, fast adsorption–desorption dynamics and good stability. Under optimal conditions, the sensitivity (IC50) of the biomimetic enzyme-linked immunosorbent assay (BELISA) method was 12.0 mg/L for trichlorfon and 30.0 mg/ L for acephate. The limit of detection (LOD, IC15) was 8.0 μg/L for trichlorfon and 12.0 μg/L for acephate. The developed method was applied to the determination of the trichlorfon and acephate in the spiked asparagus and cucumber samples with recoveries ranging from 72.1 to 92.0 % for trichlorfon and 70.0 to 85.0 % for acephate.
Keywords Molecular imprinting . Hydrophilic imprinted membrane . Biomimetic antibody . Enzyme-linked immunosorbent assay . Multi-pesticide residues . Asparagus . Cucumber
Xueyan Liu and Chen Shi contributed equally to this work. C. Shi : X. Liu : L. Song : X. Qiao : Z. Xu (*) College of Food Science and Engineering, Shandong Agricultural University, Tai’an 271018, People’s Republic of China e-mail: [email protected]
Introduction Pesticide application to grain crops increases production and maximizes resource utilization rates (Bojacá et al. 2013). Although these benefits of pesticide use could reduce global food shortages, the soil and water pollution and the human health problems caused by pesticides misuse should not be underestimated (He and Ming 2012; Susan et al. 2009). Pollution from residual pesticides, especially organophosphorus pesticides, has attracted attention in recent years. Consequently, many techniques have been used for determination of pesticide residues. Current organophosphorus pesticide determination methods include gas chromatography (Dugo et al. 2005), high performance liquid chrom atography (Seebunrueng et al. 2014), gas chromatography coupled to mass spectrometry (Sun et al. 2011), and liquid chromatography coupled to mass spectrometry (Li et al. 2013). Although these detection methods have high sensitivity and accuracy, they suffer from complex sample pretreatment processes, portability limitations because of instrument size, lengthy run times and high costs, and are not suitable for rapid, on-site testing (Suri et al. 2009). Therefore, it is necessary to establish a rapid and simple technique for pesticide residue dete
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