Development of a Highly Specific Fluoroimmunoassay for the Detection of Doxycycline Residues in Water Environmental and Animal Tissue Samples

Abstract

Doxycycline (DOX) and its metabolite residues in food and the environment pose a serious threat to human health and the ecological environment. In this work, a novel method, termed competitive fluoroimmunoassays (cFIA), based on monoclonal antibody (mAb) bio-conjugated CdSe/ZnS core–shell quantum dots (QDs), was developed for sensitive and rapid bioanalyses of DOX in natural water and commercial meats. After the optimization of the experimental conditions, 1 μg mL−1 of coating antigen and 0.5 μg mL−1 of QD-labeled mAb were used for the establishment of the cFIA. With this assay, the 50% inhibition concentration was found to be 0.35 ng mL−1 of DOX in phosphate-buffered saline samples, and the limit of detection was 0.039 ng mL−1 with minor cross-reactivity to other tetracycline members. The recoveries from natural water and commercial meats spiked with DOX concentrations of 10–600 ng mL−1 were 81.3–109.8%, and standard deviation were all below 12%. Levels measured with the QD-cFIA for thirty authentic samples were confirmed by high-performance liquid chromatography with good correlations. These results indicate that QD-cFIA is sultable for the rapid and quantitative detection of DOX residue in environmental and food samples.