M. Franek et al., ENZYME IMMUNOASSAYS FOR S-TRIAZINE HERBICIDES AND THEIR APPLICATION IN ENVIRONMENTAL AND FOOD ANALYSIS, Analytica chimica acta, 311(3), 1995, pp. 349-356
Seventeen rabbits and forty mice were immunized with immunogens prepar
ed by derivatizing atrazine and simazine and conjugating them via amin
ohexanecarboxylic acid and thiopropanecarboxylic acid substituents to
proteins. Six rabbit antisera and one murine monoclonal antibody were
incorporated into the ELISA (enzyme linked immunosorbent assay) format
using peroxidase and alkaline phosphatase tracers. Superior antisera
for atrazine, simazine and ametryn exhibited in optimized assays a 50%
binding inhibition at 0.16, 0.25 and 0.45 mu g l(-1), respectively. T
he IC50 value for the monoclonal antibody to atrazine was about 1.0 mu
g l(-1). The most sensitive antiserum Atr-(CH2)(5)-OV-3C exhibited 74
.1% cross-reactivity with propazine and 26.3% with deethylatrazine. An
tisera produced in response to the simazine-S-(CH2)(2)-determinant sho
wed a remarkable variety in s-triazine recognition selectivity. Antise
rum Sim-S-(CH2)(2)-KLH-C was extremely sensitive to ametryn, terbutryn
and prometryn with cross-reactivities based on simazine (= 100%) 2200
, 1100 and 1480%, respectively. Structural aspects of these findings a
re discussed. ELISA for atrazine was evaluated on surface water and gr
ound water samples collected from a contaminated area. The amounts of
atrazine found by ELISA were in good agreement with GC-MS analysis. St
arting data for the application of ELISA to juice and milk samples are
also presented.