Detection of Fluvalinate Residues in Beeswax and Honey Using High Performance Liquid Chromatography with Diode Array Detector

Fluvalinate is a synthetic pyrethroid widely used to control honey bee infestations caused by Varroa destructor and Acarapis woodi mites. Contamination of bee products with fluvalinate residues may pose a serious health risk to consumers and is therefore a major concern. HPLC with diode array detector (HPLC-DAD) method was developed for identification and quantification this acaricide in beeswax and honey. The method involves a modified, easy-to-perform liquid-liquid extraction of fluvalinate. As a result of selecting the optimal extracting solvent, it was found that the best extraction of the acaricide was achieved using a mixture of dichloromethane:isooctane solvents in a ratio of 2:8 (v/v). The validation characteristics of the method were satisfactory. The specificity and linearity of the method were proven. The fluvalinate recovery from beeswax and honey matrix was 85.3-96.4% and 88.7-98.9%, respectively. The limit of detection (LOD) for recovered fluvalinate from beeswax was 0.02 μg/g, from honey – 0.005 μg/g, and the limit of quantification (LOQ) was 0.05 and 0.012 μg/g, respectively. The repeatability and reproducibility of the method, expressed as relative standard deviation (RSD), were less than 5%. Also, using the proposed method, an analysis of the migration and accumulation of fluvalinate in beeswax and honey during and after the treatment of bees with fluvalinate containing strips was carried out. The study was carried out at the experimental apiary located in the Moscow region. The samples of beeswax and honey were collected both during veterinary treatment of bees (for 30 days) and after strips removal for 6 months. The data, obtained for the first time over such a long period of sample analysis, demonstrated the migration of fluvalinate from honey into beeswax and its accumulation in this matrix. The obtained results may be useful for monitoring and controlling the fluvalinate content in beekeeping products.