DEVELOPMENT OF AN ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR THE DETECTIONOF HYDROXYTRIAZINES

Hydroxytriazines are environmental decomposition products of s-triazin e herbicides. Enzyme-linked immunosorbent assays (ELISAs) in various f ormats for the detection of hydroxypropazine, hydroxyatrazine, and hyd roxysimazine were developed. methylethyl)amino]1,3,5-triazin-2-yl}amin ohexanoic acid was used as the immunizing hapten. One of the resulting ELISAs has IC50 values of 0.002, 0.004, and 0.017 mu M (0.4, 0.8, and 3.1 ppb) for hydroxypropazine, hydroxyatrazine, and hydroxysimazine, respectively. This ELISA utilizes a coated antigen format with a simaz ine-mercaptopropanoic acid hapten conjugated to conalbumin as the coat ing 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-buffere d saline. Methanol does not affect the assay up to 13%, whereas aceton itrile and acetone cause a decrease of sensitivity at 3.1%. Groundwate r 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 contain ing parts per trillion levels of hydroxytriazine contamination.