Simultaneous immunodetection of ionophore antibiotics, salinomycin and narasin, in poultry products and milk

Rabbit polyclonal antibodies were generated against the ionophore antibiotic salinomycin (SAL) as a determinant of the BSA-SAL conjugate. The homologous ELISA format was found to be preferred for similar recognition of SAL and narasin (NAR) with IC 50 values of 0.55 and 0.57 ng mL −1 , respectively. Both analytes could be determined in the range of 0.1-2.7 ng mL −1 (IC 20 -IC 80 ) with a detection limit of 0.03 ng mL −1 . To analyze matrices, individual pretreatment of samples was required. For chicken muscles, simple buffer extraction was sufficient to recover 87-110% of ionophores. Extraction with acetonitrile followed by evaporation of the solvent was best for recovering 67-108% SAL and NAR from egg homogenate. A feature of the extraction of ionophores from milk was the elimination of fat-mediated interference by organic solvation. It was found that the absence of Na + and K + ions during reconstitution of sample extracts was a key factor contributing to the increase in the average recovery of ionophores from 32% to 93%. Thanks to this special pretreatment and improved recovery, the developed immunoassay method was suitable for the analysis of ionophore antibiotics SAL and NAR in a milk matrix, which has not been previously reported. Immunoassay of ionophore antibiotics, salinomycin and narasin, was first developed for milk analysis. Good recovery was promoted by the absence of Na + -K + ions in tested samples.