Development of Improved Immunoassay and HPLC Methods for the Analysis of Chlorodiamino-s-triazine in Environmental Samples

Abstract

An enzyme-linked immunosorbent assay (ELISA) for the analysis of chlorodiamino-s-triazine (CAAT) was developed using specific rabbit polyclonal antibodies. The assay was sensitive in the submicromolar range. The method was more sensitive and specific than previously reported mouse polyclonal antibody-based ELISAs. A competitive inhibition haptenated enzyme ELISA was optimized and validated which showed that the method was accurate, precise, and reproducible. A new HPLC method was developed to resolve polar s-triazine metabolites with improvements in run time and solvent use. The ELISA method was more sensitive than the HPLC method for detecting CAAT in soil and ground water samples. Solid-phase extraction was evaluated in an attempt to further increase the sensitivity of the ELISA method. However, nonspecific interferences were observed from various types of strong cation exchange columns from different manufacturers. This made it difficult to couple a solid-phase extraction technique with the ELISA. Keyw...