The Research On Immunoassay Technology For Chemical Pestcides Such As Metolcarb

HOM, the configuration analogy with metolcarb was synthesized from the intermediate of metolcarb and P-alanine as the major materials, and it was identified by mass spectrum; next the HOM with an carboxyl end was coupled to big protein moleculors of BSA and OVA both, the coupling reaction was confirmed a success by UV-visible light spectrum scanning, the linking rate was estimated to be 33 and 17Antibody Ab01 and Ab02 was prepared from the coupled HOM-BAS, and their antiserum gradient riched 6.4 105and 1.28xl06 separately; the cross-reaction rate to the other N-methylcarbamate pestcides such as propoxur are are lower than 0.1%. It was eventually fixed for the ELISA assay conditions: coating of HOM-OVA is at 0.6(g/ml,the antibody working concentration is 20000 times the antisurem, the block of glutin is 1%, in the reaction system PBST, the icon concentration is 3.2mol/L, methanol is at 5%,the optional PH is 7.4-9. An assay curve was formed by the linking rate logit (B/Bo) and X, the logarithm of the metolcarb concentration (ug/L) as the two influencing factors, in the range of l~10000g/L, the two factor represent a apparent linear relation, the equation is as follows: Y=-9404X +1.5146, the R2 reaches 0.9939 and the sensitivity reaches 0.013ug/L, the inter-group CV reaches 2.9%, ex-group cv reaches 4.6%, the reclaiming rate from water and soil are respectively 107% and 80%.The configuration analogy methamidophosHM4 and HM5 were synthesized from the, dimethoxyphosphochlorine, an important intermediate of organophosphorus pesticide, as the main material, their structures have been improved by mass spectrum; then from HM3 and HM4 as the semi-antigens, the artificial antigens of methamidophos were coupled by the means of NHS/DCC. The visible-UV light scanning expressed that the peak curves of the coupling products and carrier proteins changed greatly, from the further study of the curves, the successful couplings of HM3-BSA, HM3-OVA. HM4-BSA and HM4-OVA are convinced. At last, the highly specific rabbit multi-antibody toward methamidophos was prepared from the former prepared HM3-BSA and HM4-BSA, so that the successful preparation of the artificial antigen of methamidophos was attested.o-isopropyloxyphenol and y- anminobutyric acid,and it has been confirmed through mass spectrum identification. Then the HOP (semi-antigen) was coupled with the big protein moleculors BSA and OVA,, its results was also improved by UV-visible spectrum scanning. The estimated coupling rate of HOP-BSA is 26 and that of HOP-OVA is 14. The obtainment of rabbit poly-antibody to propoxur. further confirmed that the artificial preparation of propoxur is a success.