Development of a Colloidal Gold Immunochromatographic Assay for the Quantitative Determination of Phenobarbital Residues in Fish and Shrimp
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Abstract:
In this study, a colloidal gold immunochromatography assay for the quantitative determination of phenobarbital residue in fish and shrimp was developed. Anti-phenobarbital monoclonal was labelled with colloidal gold nanoparticles as tracers. The assay conditions, including pH value of labelling, concentrations of antibody for labeling, concentration of the coating antigen (T line) and the goat-anti-mouse IgG (C line), as well as the sample pretreatment were optimized. By using a colloidal gold strip reader, the assay was able to achieve quantitative determination of phenobarbital. Under the optimal condition, the assay showed a limit of detection and detection range were 0.07 ng/mL and 0.08~0.94 ng/mL, respectively. The cut-off value by naked eyes was 10.0 ng/mL. The cross-reaction rates with barbital, pentobarbital and amobarbital were less than 10%, indicating a good specificity of the proposed assay. Recovery test was done by using tilapia, grass carp, perch and shrimp sample and the recoveries ranged from 73.50% to 114.17% with RSDs below 15%. The results had a good agreement with those of UPLC-MS/MS. In conclusion, the proposed assay was accurate, sensitive, convenient and rapid. It can be suitable for rapid screening of phenobarbital residues in aquatic products of fish and shrimp.
