Institutional Repository at Central Scientific Instruments Organisation (Central Scientific Instruments Organisation) · 2011 · 13 citations · 0 references
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PharmacyPharmaceutical AnalysisToxicologyAnalytical ChemistryAnalytical ToxicologyBiosensorsAssay ValidationBioanalytical Method ValidationEnzyme AssaysEnzyme-linked Immunosorbent AssayImmunoassay AnalysisImmunoassay DevelopmentAssay SensitivityAssay Sensitivity OptimizationDetection Limit AssessmentFlow Rate DependenceImmunoassay MethodologyImmunosensor-based AssaysImmunosensorsMethod Detection Limit
Quantification of atrazine in a sample was performed using a post-reactive column consisting rabbit anti-immunoglobulin G for trapping atrazine specific antibodies bound to antigen and tracer. The assay is based on a competitive enzyme immunoassay where the analyte and known amount of tracer (peroxidase labeled hapten) bind competitively to available sites of anti-atrazine \nantibodies. The absorbance signal, proportional to the amount of antigen present, is generated with the \nunbound tracer passing through the column, and monitored continuously with a spectrophotometer. Several parameters which affect the immunosensor response were evaluated, including the flow rate through the column, stability of enzyme tracer in the column, for increasing the sensitivity of the system. The immunosensor has a detection limit of 1 ng/ml with good signal precision app. 2 % in the optimum working concentration range between 1 – 100 ng/ ml of atrazine in standard water samples