Study Of Chemiluminescence Immunoassay For The Detection Of Parathion

Chemiluminescence immunoassay?CLIA?provides a highly sentitive,highly specific,easy operating,rapid,simple and economical alternative to conventional methods.This CLEIA technique has been developed and redgard as the choice for a wide variety of applications in the detection of medicine,environment monitoring,pesticide residue in agriculture production and other substance.Because of parathion high toxicity,neurological damage to human body,has been prohibited in agriculture.In order to parathion was applied in agriculture.Therefore,it demonstrated the great significance for the establishment of a rapid,simple?high throughput for the detection of parathion in agro-products.The results and main contents of the paper were listed as below:Horseradish peroxidase-H₂O₂-luminol?HRP-H₂O₂-Luminol?system and alkaline phosphatase3-?2'-spiroadamantane?-4-methoxy-4-?3'-phosphoryloxy?phenyl-1,2-dioxetane?ALP-AMPPD?system have established in CLEIA in this paper.Comparing the two system of sensitivity,stability,detection of limit?LOD?and linearity range,developing a method for trace detecting parathion in agro-products.By CLEIA procedure,the working concentrations of the coating antibody and hapten-antigen were studied based on the RLU_max and IC₅₀.For HRP-H₂O₂-Luminol system,the 5?g/L coating antibody and 3?g/L enzyme-hapten antigen were selected as HRP-H₂O₂-Luminol system working concentrations.For ALP-AMPPD system,the working concentrations of coating antibody and enzyme-hapten antigen were3?g/L and 1.5?g/L.The HRP-H₂O₂-Luminol system of CLEIA offer the possibility of improving the sensitivity of immunoassays with an IC₅₀ of 5.71?g/kg compared to that of 6.22?g/kg by the ALP-AMPPD system of CLEIA.It can be also observed that the HRP-H₂O₂-Luminol system provided a lower LOD(IC₁₀:inhibition was 10%)by comparison to the ALP-AMPPD system?0.16 and 0.26?g/kg,respectively?.In addition,the HRP-H₂O₂-Luminol system had a more wide detectable range of0.16-206.88?g/kg than that of 0.26-151.18?g/kg the ALP-AMPPD system of CLEIA.Due to the higher sensitivity,low LOD,and wide detectable range of the HRP-H₂O₂-Luminol system of CLEIA,Therefore,the established HRP-H₂O₂-Luminol system CLEIA was a more practical system than the ALP-AMPPD system for parathion.HRP-H₂O₂-Luminol-enhancer chemiluminescent?CL?system was studied in this paper.To get a CL system with high CL intensity and stability,the enhancer PIP,widely studied and applied in research,was chosen in HRP-H₂O₂-Luminol-enhancer chemiluminescent?CL?system.The effect of several factors including organic solvents,enhancers,HRP,H₂O₂ and Luminol to the chemiluminescent system were investigated.The results indicated that the concentration of DMF,HRP,Luminol,enhancer?PIP?and H₂O₂ were 10%,20?g/L,1mmol,0.5 mmol and 2 mmol,the HRP-H₂O₂-Luminol-enhancer chemiluminescent?CL?system could get the best performance in CL intensity and stability of CL-knetics.Besides,the PIP and other substance such as 4-IMP,BIP,DIOP and HIOP,which may promote to luminescence intesity in CL system,was studied in this paper.For 4-IMP and BIP,the CL intensity and the stability of CL system were decreased,which may the PIP?BIP and 4-IMP molecular structure affect the luminescence;For the DIOP,the CL intensity was enhanced but the stability of CL system wasn't improved;For the HIOP,the CL intensity?2.3×10⁶?and the stability?last to 25 min?was enhanced and prolonged.The results demonstrated the PIP and HIOP could regard as co-enhancer in HRP-H₂O₂-Luminol-enhancer chemiluminescent?CL?system.It could meet the satisfactory performance in CL intensity.The chemiluminescence enzyme immunoassay?CLEIA?method responds differently to various sample matrices because of the matrix effect.In this work,the CLEIA method was coupled with molecularly imprinted polymers?MIPs?synthesized by precipitation polymerization to study the matrix effect.The sample recoveries ranged from 72.62%to 121.89%,with a relative standard deviation?RSD?of 3.74-18.14%.The ratio of the sample matrix-matched standard curve slope rate to the solvent standard curve slope was 1.21,1.12,1.17,and 0.85 for apple,rice,orange and cabbage in samples pretreated with the mixture of PSA and C₁₈.However,the ratio of sample?apple,rice,orange,and cabbage?matrix-matched standard-MIPs curve slope rate to the solvent standard curve was 1.05,0.92,1.09,and 1.05 in samples pretreated with MIPs,respectively.The results demonstrated that the matrices of the samples greatly interfered with the detection of parathion residues by CLEIA.The MIPs bound specifically to the parathion in the samples and eliminated the matrix interference effect.Therefore,the CLEIA method have successfully applied MIPs in sample pretreatment to eliminate matrix interference effects and provided a new sensitive assay for agro-products.