Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Hydroxytriazines

Abstract

Hydroxytriazines are environmental decomposition products of s-triazine herbicides. Enzyme-linked immunosorbent assays (ELISAs) in various formats for the detection of hydroxypropazine, hydroxyatrazine, and hydroxysimazine were developed. 6-{4-Methoxy-6-[(1-methylethyl)amino]-1,3,5-triazin-2-yl}aminohexanoic acid was used as the immunizing hapten. One of the resulting ELISAs has IC50 values of 0.002, 0.004, and 0.017 μM (0.4, 0.8, and 3.1 ppb) for hydroxypropazine, hydroxyatrazine, and hydroxysimazine, respectively. This ELISA utilizes a coated antigen format with a simazine−mercaptopropanoic acid hapten conjugated to conalbumin as the coating antigen. The assay parameters are unchanged at pH values between 5.0 and 8.0 and at ionic strengths from 0.13 to 1.0 M phosphate-buffered saline. Methanol does not affect the assay up to 13%, whereas acetonitrile and acetone cause a decrease of sensitivity at 3.1%. Groundwater fortified with hydroxyatrazine was analyzed according to this method. Good correlations with spike levels were observed, making this assay potentially valuable for the screening of groundwater samples containing parts per trillion levels of hydroxytriazine contamination. Keywords: s-Triazine; hydroxytriazine; hydroxyatrazine; hydroxysimazine; hydroxypropazine; ELISA; enzyme immunoassay; herbicide metabolites

Rapid assays for environmental and biological monitoring

“…Herbicides s-Triazine herbicides are used worldwide. From a library of haptens and polyclonal and monoclonal antibodies, ELISAs for the detection of several parent s-triazines and their metabolites in natural waters, soil, urine, and foods have been developed Schneider and Hammock, 1992;Lucas et al, 1993;Kido et al, 1995;Jaeger et al, 1995). ELISAs were devised for several arylurea herbicides (diuron, monuron, and linuron) (Schneider et al, 1994;Kam et al, 1994).…”

Development of an ELISA for the Organophosphorus Insecticide Isofenphos

“…The assay conditions were the same as that described above. The effect of ionic strength on ELISA performance was studied using different concentrations of PBST (10,90,190 and 390 mM phosphate resulting in the final concentration of 10, 50, 100 and 200 mM, respectively) to dilute the antisera and 10% methanol-PBS to dissolve the pesticide. The procedure for the competition assay was as follows.…”

An approach to the construction of an immunoarray for differentiating and quantitating cross reacting analytes

“…The polyclonal antibodies 194, 841 and 842 were produced by Harrison et al [9]. The polyclonal antibody 2652 was produced by Lucas [18], the polyclonal 4653 is described in [30]. Five of the triazine herbicide derivatives were synthesized by Goodrow et al [31], one by Muldoon et al [20].…”

“…MAb KIF4 prefers analytes substituted in the R1.position (terbutryn, prometryn, prometon) but cross reacts with some other analytes as well [15]. Pab 4653 reacts almost exclusively with hydroxyatrazine, but at higher concentrations prometon can be also detecte,d [30]. Pab 2652 detects only Initially we only compared antibody responses to identical concentrations (in ppb) of the eight analytes.…”