Dipstick Format of an Improved ELISA for On-Site Atrazine Monitoring in Water in Pakistan
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 2030C 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 g/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 g/L. However, dipsticks were superior, fast, and cost-effective.
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Document Type: Research Article
Affiliations: Nuclear Institute for Agriculture and Biology, PO Box 128, Faisalabad, Pakistan.
Publication date: 01 January 2010
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