Click reaction triggered turn-on fluorescence strategy for highly sensitive and selective determination of steroid hormones in food samples

Abstract The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction has becoming noticeable in the field of analytical chemistry. Mild reaction conditions, simple operation, high efficiency, and good regioselectivity make this classical click reaction a perfect strategy for chemical derivatization. Herein, we proposed a promising click fluorescent labeling method with high selectivity for the determination of five steroid hormones in food samples. The labeling strategy depends on the reaction between 3-Azido-7-hydroxycoumarin and the alkynyl group of steroid hormones, which shows a turn-on fluorescence response in the presence of copper (I). The formed fluorescent products were detected by HPLC-FLD in the range of 395 to 470 nm. Under the optimized conditions, the proposed method presented excellent performance with good linearity (R2 ≥ 0.9998) and low detection limit (1.8-7.3 μg L-1). Further, satisfactory recoveries were obtained to be 82-107% in spiked meats with relative standard deviations (RSDs) ≤ 5.7%. Finally, the established method was successfully applied for the determination of steroid hormones in meat, indicating the potential prospect of the click reaction in chemical derivatization.