Differential detection of N-heterocyclic compounds and their N-methylated derivatives by immunoanalysis

Competitive enzyme-linked immunosorbent assay (ELISA) systems are proposed for the indirect monitoring of formaldehyde by the parallel detection of its N-methylated precursors and the corresponding demethylated compounds. As an example for such immunoanalytical differentiation between an N-heterocyclic compound and its N-methylated derivative, the quantitative detection of the systemic triazole fungicide, myclobutanil, is discussed. Antibodies recognizing the non-zwitterionic structure of 2-(4-chlorophenyl)-2-[(1,2,4-triazol-1-yl)-methyl]-hexanonitril e (myclobutanil) showed only minor binding to corresponding N-alkylated derivatives of myclobutanil. And vice versa, literature data indicate that antibodies raised against the pyridilium ionic structure of the herbicide paraquat, displayed only mediocre reactivity towards the corresponding dealkylated derivatives. Thus, both experimental and literature data suggest that immunoanalytical methods for differential detection of N-methylated heterocycles (potentially including formaldehyde precursors) and their non-methylated counterparts are possible to develop.