The Determination of Six Ionophore Coccidiostats in Feed by Liquid Chromatography with Postcolumn Derivatisation and Spectrofotometric/Fluorescence Detection

The control of levels of anticoccidial feed additives in targeted feeds plays an important role in the assurance of efficiency of animal treatment, prevention of drug resistance, and food safety. The robust and labour-efficient method for the simultaneous determination of six ionophore coccidiostats (lasalocid, maduramicin, monensin, narasin, salinomycin, and semduramicin) in targeted feed has been developed. Properly grinded and homogenized feed sample was spiked with internal standard (monesin methyl ester) and extracted with methanol. The extract was analysed with reversed phase HPLC without any further purification. The separation of the analytes with conventional C18 and core-shell columns was compared. Lasalocid was analysed with fluorescence detection, whereas other ionophores were detected with UV-Vis detector after derivatisation with vanillin in the presence of sulfuric acid. Fortified samples and targeted feeds at authorized levels were used for method validation. Recovery was in the range of 85-110%, depending on the analyte. The within-laboratory reproducibility did not exceed the target value from Horwitz equation. The results of the proficiency tests (z-scores in the range of -1.0 to 1.9) confirmed the reliability of the developed protocol.