Development of an Enzyme-Linked Immunosorbent Assay for Residue Analysis of the Insecticide Emamectin Benzoate in Agricultural Products

Abstract

A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) for the analysis of emamectin residues in agricultural products was developed using a prepared mouse monoclonal antibody. The working range was 0.3-3.0 ng/mL, and the 50% inhibition concentration (IC(50)) was 1.0 ng/mL. The assay was sufficiently sensitive for analysis of the maximum residue limits in agricultural products in Japan (>0.1 microg/g). Emamectin residues contain the following metabolites: the 4''-epi-amino analogue, the 4''-epi-(N-formyl)amino analogue, the 4''-epi-(N-formyl-N-methyl)amino analogue, and the 8,9-Z isomer. The dc-ELISA reacted with these compounds at ratios of 113, 55, 38, and 9.1% of the IC(50) value of emamectin benzoate. Seven kinds of vegetables were spiked with emamectin benzoate at concentrations of 15-300 ng/g, and the recoveries were 91-117% in the dc-ELISA. The dc-ELISA results agreed reasonably well with results obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using spiked samples and actual (incurred) samples. The results indicate that the dc-ELISA was useful for the analysis of emamectin benzoate residues in agricultural products.

Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran

“…Designing the hapten is one of the most important steps in preparation of a MoAb which reacts with the target pesticides [19,21–25]. Four compounds among the neonicotinoid insecticides shown in Figure 1 have a six-membered pyridine ring in their structure.…”

Quantitative analysis of fungicide azoxystrobin in agricultural samples with rapid, simple and reliable monoclonal immunoassay

“…acetone, acetonitrile and methanol, are essential to quantitative extraction of azoxystrobin from agricultural samples when chromatographic methods are used (Fenoll et al, 2007;Hernández et al, 2006;Leandro et al, 2007;Štajnbaher & Zupančič-Kralj, 2003;Teixeira et al, 2004). In immunochemical methods, methanol is the most easy-to-use organic solvent because of relatively low impact for denaturation of proteins, such as HRP and antibody, which are used in ELISA (Kondo et al, 2009(Kondo et al, , 2012Watanabe et al, 2006Watanabe et al, , 2011. Since sample solutions always contain methanol as an extraction solvent, the kit-based dc-ELISA was designed to perform azoxystrobin determination under 10% methanol.…”

Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for the Organophosphorus Insecticide O-Ethyl O-4-Nitrophenyl Phenylphosphonothioate (EPN)

“…Table 4 shows the cross-reactivities that were determined by both the indirect and direct assays, Comparison between MAb-Based and PAb-Based ELISA. It has been over three decades since immunoassays began to be developed for detecting pesticides, but studies in this field are still actively carried out (24)(25)(26)(27). It is now certain that most widely used pesticides can be detected by immunoassay methods.…”