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Application of an Enzyme-Linked Immunosorbent Assay for the Analysis of Paraquat in Human-Exposure Samples

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Abstract

Paraquat is a toxic quaternary ammonium compound used as an herbicide around the world. Easy, fast, and inexpensive but sensitive methods are needed to study the effects of long-term, low-level exposure of paraquat on human health. An enzyme-linked immunosorbent assay (ELISA) was used for quantification of paraquat in urine and air-filter samples collected in a human-exposure study among farm workers in Costa Rica. A sample pretreatment consisted of removal of interfering substances using solid-phase extraction resin columns. The precision and accuracy of the method were tested using duplicate spiked urine samples. The correlation between results for blind samples obtained using ELISA and liquid chromatography–mass spectrometry was significant (R2 = 0.945 and 0.906 for spiked and field samples, respectively). With an LOQ of 2 ng mL−1, this ELISA method was able to distinguish the exposed from the nonexposed farm workers. For the air-filter analysis, paraquat was extracted by 9M H2SO4 at 60°C for 12 hours, and the results obtained by ELISA showed good correlation (R2 = 0.918) with the spectrophotometric (256 nm) measurements. Paraquat in acid-stabilized urine samples was very stable, and no significant losses were detected during a 3-month storage at room temperature, at 4°C, or at −20°C.

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Acknowledgments

The authors thank Dr. B. Woollen from Syngenta for carrying out the LC-MS analysis for the set of urine samples used in quality assessment for the ELISA method and Dr. Marvin Goodrow for the synthesis of the paraquat haptens. We acknowledge and thank the members of the science advisory panel, Drs. David Christiani (chair), Richard Fenske, Bertold Hock, Rogelio Perez-Padilla, and Mark Segal for careful and critical review of the scientific aspects of the study. This work was supported in part by grants from the National Institute of Environmental Health Sciences (Grant No. 5 P42 ES04699); the NIEHS Center for Environmental Health Sciences (Grant No. P30 ES05707); the NIOSH Center for Agricultural Disease and Research, Education, and Prevention (Grant No. 1 U50 OH07550); and Syngenta Research Agreement No. 002994.

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Authors and Affiliations

  1. Department of Entomology and the University of California Davis Cancer Center, University of California, Davis, California, 95616, USA

    M. E. Koivunen, S. J. Gee, E. -K. Park & B. D. Hammock

  2. Department of Epidemiology and Preventive Medicine, University of California, Davis, California, 95616, USA

    K. Lee & M. B. Schenker

Authors
  1. M. E. Koivunen
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  2. S. J. Gee
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  3. E. -K. Park
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  4. K. Lee
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  5. M. B. Schenker
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  6. B. D. Hammock
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Corresponding author

Correspondence to B. D. Hammock.

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Koivunen, M.E., Gee, S.J., Park, E.K. et al. Application of an Enzyme-Linked Immunosorbent Assay for the Analysis of Paraquat in Human-Exposure Samples. Arch Environ Contam Toxicol 48, 184–190 (2005). https://doi.org/10.1007/s00244-003-0251-x

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  • DOI: https://doi.org/10.1007/s00244-003-0251-x

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