Development of a Fluorescence Polarization Immunoassay for the Quantification of Tylosin in Honey

A fluorescence polarization immunoassay method has been developed for the determination of antibiotic tylosin in honey. For this purpose, rabbits were immunized with a conjugate of tylosin and bovine serum albumin, and antisera were obtained. The conditions for the interaction of antisera in a competitive format with tylosin in the sample and with tylosin labeled with fluorescein were optimized. The developed method is characterized by a limit of detection for tylosin of 34.7 ng/mL and an analytical range of 65.2 to 564.0 ng/mL. High reproducibility of measurements using a portable detector was demonstrated, with relative standard deviation (RSD) values of 1.5-2.5%. The method was validated using honey samples, showing recovery ranging from 98 to 103.8%. An analysis time, excluding sample preparation, is 10 min.