Hapten synthesis for enzyme-linked immunoassay of the insecticide triazophos.
Two haptens of the insecticide triazophos (O,O-diethyl O-[1-phenyl-1H-1,2,4-triazol-3-yl] phosphorothioate) were synthesized by introducing appropriate spacers in the O-ethyl site of the analyte molecular structure. First, thiophosphoryl chloride (PSCl(3)) reacts with methanol at low temperature to give O-ethyl dichlorothiophosphate. After reacting with 1-phenyl-3H-1,2,4-triazol, the O-ethyl dichlorothiophosphate was transformed into the intermediate O-ethyl O-(1-phenyl-1H-1,2,4-triazol-3-yl) chlorothiophosphate. Then the intermediate reacts with 4-aminobutyric acid and 6-aminobutyric acid to produce hapten I and hapten II, respectively. The molecule structures of the two haptens were identified by (1)H nuclear magnetic resonance spectrum and mass spectrum. An enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody was also developed to evaluate the two haptens. Results showed that the monoclonal antibodies with high titers were obtained after immunizing with protein conjugates of these haptens and that the immunoassay has high affinity and specificity to triazophos. These results suggested that the haptens were synthesized successfully and could be used for immunoassay for the rapid screening and sensitive determination of this insecticide.
Gui
Wen Jun
WJ
Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou 310029, China.
Jin
Ren Yao
RY
Chen
Ze Li
ZL
Cheng
Jing Li
JL
Zhu
Guo Nian
GN
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D001201:Ascites; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006241:Haptens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007306:Insecticides / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D063086:Organothiophosphates / Q000032:analysis* / Q000737:chemistry; D012015:Reference Standards; D014230:Triazoles / Q000032:analysis* / Q000737:chemistry
D000911:Antibodies, Monoclonal; D006241:Haptens; D007306:Insecticides; D063086:Organothiophosphates; D014230:Triazoles; C012584:triazophos