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United States Department of Agriculture

Agricultural Research Service

Research Project: INTERVENTIONS TO REDUCE EPIZOOTIC PATHOGENIC BACTERIA IN SWINE AND CATTLE

Location: Food and Feed Safety Research

Title: A simple, rapid, and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples

Authors
item Xu, Zhen-Lin -
item Wang, Qiang -
item Lei, Hong-Tao -
item Eremin, Sergei -
item Shen, Yu-Dong -
item Wang, Hong -
item Beier, Ross
item Yang, Jin-Yi -
item Maksimova, Ksenia -
item Sun, Yuan-Ming -

Submitted to: Analytica Chimica Acta
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 30, 2011
Publication Date: October 6, 2011
Citation: Xu, Z., Wang, Q., Lei, H., Eremin, S.A., Shen, Y., Wang, H., Beier, R.C., Yang, J., Maksimova, K.A., Sun, Y. 2011. A simple, rapid, and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples. Analytica Chimica Acta. 708:123-129.

Interpretive Summary: Organophosphorus pesticides (OPs) have been widely used in agricultural and domestic applications for more than six decades. However, OP intoxication can occur in humans and animals. We have developed antibodies that recognize a broad group of commonly used OPs. Atibodies are substances that are produced by the immune system in response to foreign substances which enter the body. Once the antibodies to a foreign substance are isolated, they can be used in a method to detect the presence of that foreign substance. The antibodies that we isolated may be used in an easy-to-use test called a fluorescent polarization immunoassay, or FPIA, for the detection of OPs. A fluorescent polarization immunoassay is based on interactions between a fluorescein-labeled antigen (called a tracer) and a specific antibody. This comination allows the determination of specific analytes within a very short period of time, and the immunoassay can be accomplished in a single reaction solution without separation or washing steps. In this work, we evaluated the FPIA under different situations to obtain the best sensitivity. The designed method is capable of simultaneously determining 12 different OPs in 10 minutes. The evaluation conducted here demonstrated that this assay is ideally suited as an initial screening method for OP residues in vegetable and environmental water samples.

Technical Abstract: A simple, rapid, and high-throughput fluorescent polarization immunoassay (FPIA) for simultaneous determination of organophosphorus pesticides (OPs) was developed. Three haptens were labeled with a fluorescein probe and used as tracers to develop a homogenous FPIA using a broad-specificity monoclonal antibody for OPs. The FPIA can detect twelve OPs simultaneously with a limit of detection below 80 ng mL-1. The total time required for the measurement of 42 samples in duplicate was less than 10 min. The recovery from spiked vegetable and environmental samples ranged from 84.8% to 124.0%, with the coefficient of variation ranging from 5.6% to 15.0%. The correlation coefficient between FPIA and HPLC-MS/MS analysis was 0.9645. The proposed FPIA exhibited good accuracy and reproducibility and is suitable for rapid and high-throughput testing for OP contamination in vegetable and environmental water samples with high-efficiency and low cost.

Last Modified: 4/23/2014
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