Development of an enzyme-linked immunosorbent assay (ELISA) for the herbicide propanil

Vitalija Krikunova, Gyöngyvér Hegedus, Hong My Le, Loudmila Jouravleva, Sergei Eremin, Marco Natangelo, Emilio Benfenati, András Székács

Research output: Contribution to journalArticle


Although the extensively used postemergence herbicide propanil itself is of low acute toxicity in mammals, it raises environmental concerns due to its effect on aquatic organisms and other adverse impacts. Therefore, in order to obtain a rapid analytical method for this pesticide, an indirect enzyme-linked immunosorbent assay (ELISA) has been developed. Antibodies obtained against a conjugate of 3,4-dichloroaniline coupled to succinylated proteins were tested in hapten-homologous and heterologous indirect ELISA formats using various N-(dichlorophenyl)-succinamic acid derivatives conjugated to carrier proteins as coating antigens. Titers in ELISA were found to be significantly affected by the type and quantity of coating antigen. One of the optimized systems using N-(2,4-dichlorophenyl)-succinamic acid and N-(3,5-dichlorophenyl)-succinamic acid conjugated to ovalbumin allowed serum dilution of 1:10,000 and IC50 values of 2.2 and 2.7 ng/mL for propanil, respectively. The limit of detection (LOD) of the immunoassays is 0.2 ng/mL. Other optimized ELISA systems based on different dichloroaniline-based coating antigens also offered similar sensitivities. The ELISA systems appeared to tolerate methanol and ethanol upto 5% concentration. For confirmatory purposes, the ELISA protocol was compared with a highly sensitive gas chromatographic method coupled with mass spectrometric detection (GC-MS). Spiked propanil content was detected both by ELISA and GC-MS in methanolic rice extract. Detection sensitivities of the two analytical systems appeared to closely correlate with each other in the range of 10-90 ng/mL (0.02-0.18 μg/g), indicating the utility of the immunoanalytical method in detecting propanil content in rice, the main commodity propanil is being applied on.

Original languageEnglish
Pages (from-to)865-878
Number of pages14
JournalInternational Journal of Environmental Analytical Chemistry
Issue number11-12
Publication statusPublished - Dec 2002


  • Enzyme immunoassay
  • Optimization
  • Propanil
  • Sensitivity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Environmental Science(all)
  • Environmental Chemistry

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