Abstract:[Objective]This study aims to establish a rapid immunoassay method for the quantitative detection of eugenol.[Method]The eugenol monoclonal antibody was conjugated to the surface of quantum dots by the 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxy-succinamide (NHS) (EDC-NHS) coupling method, with quantum dots as fluorescent markers. Bovine serum albumin (BSA) was used for blocking, and fluorescence microscopy, agarose gel electrophoresis, and a microplate reader were used for identification. Furthermore, the established method was compared with high performance liquid chromatography (HPLC).[Result]The optimal synthesis conditions were as follows: quantum dot activation time of 30 min, conjugation pH with 8.0, antibody addition of 21 μL, and conjugation incubation time of 3.0 h. The optimal buffer system was as follows: carbonate buffered solution (CBS) with the coating buffer solution of 0.05 mol/L and pH 9.6, phosphate buffered solution (PBS) with the quantum dots-monoclonal antibody (QDs-mAb) concentration of 0.01 mol/L and pH 7.4. The quantum dots-fluorescence immunoassay (QDs-FIA) method was established under optimized conditions, with the linear equation of y=13.098ln x+9.625 8, R2=0.993 1. The IC20, IC50, and IC80 were 2.21, 21.81, and 215.49 ng/mL, respectively. Recovery test were performed on water samples and bass meat samples, with recovery rate ranging from 96.32% to 103.01% for water samples and 94.16%~102.28% for bass meat samples, and the coefficients of variation were less than 7%.[Conclusion]The established QDs-FIA method has good sensitivity and accuracy, which can provide a theoretical basis for the development of fluorescent assay kit in the future.
