New incorporation of the S9 metabolizing system into methods for detecting acetylcholinesterase inhibition

摘要: The activity of individual biotransformation products cannot be measured in multicomponent mixtures by the current status-quo assays. A prior separation and tedious isolation compounds is required, often addition, a concentration step into solvent suitable for cell-/enzyme-based assay. Hence, metabolizing S9 system, mimicking complex reactions liver, was aimed to integrated two orthogonal methods analysis acetylcholinesterase (AChE) inhibition. For microtiter plate assay method, evaluation generated fluorescence signal impaired incorporated system. In contrast, metabolic activator (S9 mixture) successfully high-performance thin-layer chromatography (HPTLC) method. As proof principle, four reference AChE inhibitors were studied samples with without activation. addition neurotoxic carbamate eserine organophosphate insecticides chlorpyrifos, quinalphos parathion, tris(nonylphenyl) phosphite nonylphenol, both originating from food contact materials, tested but also packaging migrate extract. method comparison benchmarking pointed multifold advantages using this newly developed bioanalytical tool assessment neurotoxins samples. sensitive HPTLC-S9-AChE allowed detection chemicals activation, at levels consistent threshold toxicological concern organophosphates carbamates. This new on surface metabolism system can applied other toxicities