Rapid and sensitive determination of clenbuterol in porcine muscle and swine urine using a fluorescent probe

Abstract

The feed additive Clenbuterol hydrochloric acid (CLB) is non-fluorescent, thus it is difficult to quantify through direct fluorescent method. Palmatine (PAL) can react with cucurbit[7]uril (CB[7]) to form stable complexes as a fluorescent probe. Significant quenching of the fluorescence intensity of the CB[7]–PAL complex was observed with the addition of CLB. Based on the significant quenching of the supramolecular complex fluorescence intensity, a novel spectrofluorimetric method with high convenience, selectivity and sensitivity was developed for the determination of CLB. The fluorescence quenching values (ΔF) showed good linear relationship with CLB concentrations from 0.011μgmL−1 to 4.2μgmL−1 with a detection limit 0.004μgmL−1. In this research, an ultrasound treatment replaced the former time-consuming shake method to form stable complexes. The proposed spectrofluorimetric method had been successfully applied to the determination of CLB in porcine muscle and swine urine with good precision and accuracy. The competing reaction and the supramolecular interaction mechanisms between the CLB and PAL as they fight for occupancy of the CB[7] cavity were studied using spectrofluorimetry, 1H NMR, and molecular modeling calculations. Interestingly, results indicate that two stable CB[7]–CLB complexes were formed.