| Nonylphenol(NP) is considered as one of the confirmed environmental endocrine disruptors.The polluting condition in environment is severe enough to be monitored and evaluated since it has been taken as an important part in environmental surveillance and control.Although the accurate results can be obtained by advanced apparatus,but it's time-consuming,laboring and costing.The main purpose of this research is the rapid detection of biological technology for NP.Neighbor active hydrogen of "-OH-"in NP was replaced by organic chemical methods and coupled to amino group in the protein carrier,and then,the NP-protein conjugate was formed.The combining rate turns to be 18:1 and 12:1 with the aid of UV-Vis Spectrophotometer for the detection of the absorption values.The poly-antibody was prepared by the use of NP-protein conjugate and the titer was determined as 1.78125.The optimum condition for the sample detection was determined by chessboard titration experiment.The lowest detection limit is determined as 8ng/ml by detection of a series of standards and the plotting of the standard curve.Meanwhile,the specificity is good.Nano-colloidal gold was prepared by heating and mixing with sodium citrate reduction of the acid chloride.The diameter size of the particle and the even degree were obtained by UV-Vis Spectrophotometer scanning.The diameter size of the particle is approximately 19nm and the distribution is fairly even by observation of the field emission TEM.By exploration of the optimum condition,the optimum pH is 7.6 for the effective combination of the NP poly-antibody and the colloidal gold was determined.The optimum addition of the protein is 24μg by adding colloidal gold per milliliter.Based on the above condition mentioned,the immune colloidal gold was prepared and the progress was monitored.The conjunct is immunocompetent and confirmed by dot immunogold filtration assay.With comparison,the release pad of the colloidal gold is glass film with the model SB06 made by Jinbiao biotec co,Shanghai.The concentration of the NP antibody on the film is 48μg/ml.The material of the chromatography is pyroxylin film made by Millipore co.with the model HFB1350220,in which the sheep anti-rabbit IgG coated as control line,the concentration of sheep anti-rabbit IgG dilutionin 1:10,the NP-OVA coated at 0.52mg/ml as the test line.The sensitivity is(4.4±2.5)μg/ml by 10 NP samples detection using immunochromatography test paper.The test paper can be sealedly stored for 6 months with the adding of desiccant in 4℃with the original steady sensitivity.Clear bands are appearing within 10min.Requirement of rapid detection can be reached which the testing time is short, good stability.NP pollution incidents can be achieved by its rapid and qualitative detection.Compared with the method of GC-MS,both ELISA and GICA were not necessary to deal the sample before detection,then,it is timesaving and more economical.The sensitiveness of ELISA reached to the level of the same method in foreign laboratory and the sensitiveness of GICA got close the level of immunobiochip in our country.The sensitiveness is lower than GC-MS using the mentioned two methods,but for achieving the purpose of rapid detection it is have it's value,...
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