Abstract
This study describes the development of a reproducible and label-free surface plasmon resonance (SPR) immunosensor and its application in the detection of harmful enrofloxacin (ENRO) in animal-derived foods. The experimental parameters for the immunosensor construction and regeneration, including the pH value (4.5), concentration for coating ENRO-ovalbumin conjugate (ENRO-OVA) (100 μg·mL-1), concentration of anti-ENRO antibody (80 nM) and regeneration solution (0.1 mol·L-1 HCl) were evaluated in detail. With the optimized parameters, the proposed SPR immunosensor obtained a good linear response to ENRO with high sensitivity (IC50: 3.8 ng·mL-1) and low detection limit (IC15: 1.2 ng·mL-1). The proposed SPR immunosensor was further validated to have favorable performances for ENRO residue detection in typical animal-derived foods after a simple matrix pretreatment procedure, as well as acceptable accuracy (recovery: 84.3-96.6%), precision (relative standard deviation (n = 3): 1.8-4.6%), and sensitivity (IC15 ≤ 8.4 ng·mL-1). Each SPR chip for analysis can be reused at least 100 times with good stability and the analysis cycle containing the steps of sample uploading/chip regeneration/baseline recovery can be completed within 6 min (one cycle) and auto-operated by a predetermined program. These results demonstrated that the proposed SPR immunosensor provided an effective strategy for accurate, sensitive, and rapid detection for ENRO residue, which has great potential for routine analysis of large numbers of samples for measuring different types of compounds.