Determination of aflatoxin-B1 in poultry feed and its components employing enzyme-linked immunosorbent assay (ELISA)

A highly sensitive enzyme immunoassay was standardized for aflatoxin B₁ determination in poultry feed and its components using Riedel-de-Haen, ELISA Systems_. A microtitration plate method was optimized using anti-aflatoxin B₁ antibodies and peroxidase – aflatoxin B₁ conjugate, based on competitive enzyme immunoassay principle. Standards of concentrations of 5, 10, 20, 50, 100, 500?ng/L aflatoxin B₁, prepared in phosphate-buffered saline, were used. Standard curves showed that as the concentration of antigen decreased, absorbance values increased. Fifty percent inhibition was observed at 37?ng/L. Regression analysis showed that log concentration was inversely related to %B/B₀, with a highly significant negative correlation (?0.980). The lowest detection limit for aflatoxin B₁ was 5?ng/L. Using this standardized ELISA, aflatoxin B₁ was detected in most of the commercially available poultry feed samples and their components.

The data suggest that this test is suitable for the accurate determination of aflatoxin B₁ concentrations in poultry feed and its components.     

Development of ELISA technique for the analysis of atrazine residues in water

A highly sensitive enzyme immunoassay is described for the detection of atrazine residues in water. Atrazine derivative was conjugated to Bovine Serum Albumin (BSA) to obtain an immunizing antigen and to Horseradish Peroxidase enzyme (POD) to obtain a marker for immunoassay. The formation of these conjugations was confirmed by UV spectroscopy as well as by gel-electrophoresis. Polyclonal antibodies were raised in rabbits by immunization with an atrazine-BSA conjugate containing 29 atrazine residues per BSA molecule. An ELISA on microtitration plates was optimized with peroxidase-atrazine conjugate. The middle of the test (50% B/Bo) was found to be at 90 ng/l, which is well below the maximum concentration permitted by the EC guidelines for drinking water. Detection limits for atrazine of about 1 ng/l could be reached. The assay did not require concentration or cleanup steps for drinking or ground water samples. Validation experiments showed good accuracy and precision. No cross-reactivities were shown by other s-triazines like terbutryn, ametryn, terbuthylazine, des-isopropylatrazine, and de-ethylatrazine except hydroxyatrazine. The latter was present at very low levels that can be calibrated/standardized before analysis or it may be considered as leftover residues of atrazine. Based on these results, it is suggested that this test can be applied to obtain fairly accurate results for atrazine concentration in water samples from different sources.