Dipstick format of an improved ELISA for on-site atrazine monitoring in water in Pakistan

J AOAC Int. 2010 Jan-Feb;93(1):19-27.

Abstract

A dipstick format was developed for on-site atrazine monitoring in water samples of different origins. It was derived from an in-house-developed ELISA based on polyclonal antibodies that also cross-react with hydroxyatrazine (30%) and terbuthylazine (17%). Test reagents were evaluated for temperature and pH stabilities and rapidity for field applications. Reagents performed well within a temperature range of 20-30 degrees C and were tolerant to alkaline pH (up to 8.5) of the assay buffering system. Tracer incubation time could be reduced to 40 min. Bovine serum albumin addition (1%) in the assay buffer improved assay performance, giving 50% B/B0 (IC50) of 65 ng/L and the lowest LOD of 2 ng/L at 90% B/B0 (IC10). The dipstick ELISA format was standardized on a membrane support. Nylon membrane, positively charged, was superior to PVDF for qualitative or semiquantitative analysis regarding color intensity and stability. Tracer incubation time was further reduced to 30 min with a lowest LOD of 0.1 microg/L. For real sample screening with dipsticks, acceptable results were obtained for water. Significant correlation was found between dipstick and plate ELISA results. Validation using GC with a nitrogen-phosphorus detector and HPLC indicated that dipstick signals in aged water samples, which were mainly due to hydroxyatrazine, were significantly above European Commission regulations of 0.1 microg/L. However, dipsticks were superior, fast, and cost-effective.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Atrazine / analysis*
  • Atrazine / toxicity
  • Cattle
  • Chromatography, Gas
  • Chromatography, High Pressure Liquid
  • Environmental Monitoring
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Enzyme-Linked Immunosorbent Assay / standards
  • Herbicides / analysis*
  • Herbicides / toxicity
  • Humans
  • Hydrogen-Ion Concentration
  • Pakistan
  • Reproducibility of Results
  • Serum Albumin, Bovine
  • Temperature
  • Water Pollutants, Chemical / analysis*
  • Water Pollutants, Chemical / toxicity
  • Water Supply / analysis*

Substances

  • Herbicides
  • Water Pollutants, Chemical
  • Serum Albumin, Bovine
  • Atrazine