Development Of Technique For Rapid And Sensitive Detection Of Acenaphthylene In Water Based On Enhanced ELISA

As ubiquitous environmental contaminants,polycyclic aromatic hydrocarbon(PAHs)can invade the human body through the skin,respiratory and digestive system.PAHs have genetic toxicity and embryo toxicity that can disrupt the immune system to interfere with fetal development.Acenaphthylene(ACE)as a kind of PAHs,was listed as one of the priority pollutants in water environment by U.S Environmental Protection Agency(EPA).Because PAHs are a serious threat to human health,it is of particular importance to develope a rapid and sensitive dtection for PAHs.At present,immunoassay has the advantages of high sensitivity,selectivity and specificity which is unparalleled by conventional assays and it can be utilized as a low cost and efficient detection technology for PHAs.In this study,the complete antigens of ACE were prepared by activated ester method.The BSA and OVA were used as the carrier protein to prepare immune antigen(c BSA-ACE)and detect antigen(OVA-ACE).New Zealand white rabbits were subcutaneous immunized by c BSA-ACE to produce Pc Ab.Rrapid and sensitive detection assays for ACE were developed based on the purified Pc Ab.A indirect competitive ELISA(ic-ELISA)was established and optimized.The detect time of the ic-ELISA was 5 h 10 min and the regression equation was y =20.897x-65.861 with correlation coefficient R2=0.9946.The sensitivity of the ic-ELISA was 4.26 ng/m L and the linear range was from 12.8 ng/m L to 9.5 ?g/m L.The recoveries were from 99.42% to 100.73% for the added different concentration of ACE standard solution to water.The ic-ELISA based on nanomagnetic beads(MBs-ic-ELISA)was developed and optimized.The detect time of MBs-ic-ELISA was 38 min and the regression equation of this assay was y = 23.642x-69.819 with correlation coefficient R2 =0.9915.The sensitivity of the MBs-ic-ELISA was 2.37 ng/m L and the linear range was from 3.9 ng/m L to 3.5 ?g/m L.The recovery rates were from 98.73% to 103.4%for the added different concentration of ACE standard solution to water.The ic-ELISA based on MBs and biotin and streptavidin(MBs-BS-ic-ELISA)was established and optimized.Only 22 min was needed to finish the procedure of this assay.The regression equation of MBs-BS-ic-ELISA was y =16.509x-26.761 with correlation coefficient R2=0.9998.The sensitivity of this assay was 0.16 ng/m L and the linear range was from 0.68 ng/m L to 2.93 ?g/m L.The average recoveries of ACE in tap water,mineral water and lake water samples were 102.1%,101.1% and103.8%,respectively.Five water samples from different areas(Changchun,Beijing,Changbai mountain,Sanmenxia,laboratory tap water)were collected to evaluate the application of the ic-ELISA,MBs-ic-ELISA and MBs-BS-ic-ELISA.The concentration of ACE was 14.89 ng/m L in Sanmenxia water sample detected by the ic-ELISA.And ACE was not detected in other water samples.The concentration of ACE in five water samples(Changchun,Beijing,Changbai mountain,Sanmenxia,laboratory tap water)detected by the MBs-ic-ELISA were 0 ng/m L,3.74 ng/m L,0 ng/m L,13.92 ng/m L,0ng/m L.And the results of MBs-BS-ic-ELISA were 1.07 ng/m L,3.85 ng/m L,0 ng/m L,14.96 ng/m L,0 ng/m L.In this paper,the Pc Ab of ACE was produced and the ic-ELISA,MBs-ic-ELISA,MBs-BS-ic-ELISA were developed respectively.The detection time and sensitivity of MBs-ic-ELISA and MBs-BS-ic-ELISA were improved in different degrees to meet various demand detection.