Development and Validation of a Multiclass, Multiresidue Method for Veterinary Drug Analysis in Infant Formula and Related Ingredients Using UHPLC-MS/MS

A multiclass, multiresidue method based on ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) has been developed and validated for the analysis of around 150 veterinary drugs in infant formula and related dairy ingredients. The included analytes belong to the following veterinary drug classes: anthelmintics, antibiotics (aminoglycoside, amphenicols, beta-lactams-penicillins and cephalosporins, lincosamides, macrolides, quinolones, sulfonamides, tetracyclines, and others), antimicrobial growth promoters, antiprotozoals, beta-agonists, coccidiostats, dyes, pesticides, and tranquilizers. The sample preparation procedure involves dispersing the sample in 0.05 M EDTA solution in water, followed by extraction with 0.1% formic acid in acetonitrile, drying down an aliquot of the extract, and reconstituting it in a water-acetonitrile mixture. The analyte detection, identification, and quantitation are performed by UHPLC-MS/MS using positive electrospray ionization mode. The method was validated in infant formula powder, whole milk powder, and whey protein isolate, typically achieving limits of quantitation (meeting acceptable recovery and precision validation criteria) at 1-10 ng/g.