Detection of selected corticosteroids and anabolic steroids in calf milk replacers by liquid chromatography–electrospray ionisation – Tandem mass spectrometry

• Powdered milk is commonly used for the daily feeding of calves.
• Drug inclusion in milk replacers is forbidden in the EU and has to be reduced in USA.
• An analytical method was fully validated for 5 corticosteroids and 8 anabolic steroids.
• In order to check the method, 50 samples were analysed.
• β-nandrolone and prednisolone were detected in addition to endogenous steroids.

The use of corticosteroids and anabolic steroids in food producing animals is regulated or banned in the European Union (EU). However, their use as growth promoters cannot be excluded. Milk replacers, considered by EU legislation as feeds, may be a good way of administration of these compounds. In order to improve the control of growth promoter utilization in animal husbandry and preventing possible consequences to animal welfare, we developed a method for multiresidue analysis of prednisolone, prednisone, dexamethasone, cortisone, cortisol, 17α- and 17β-boldenone and their precursor androstadienedione (ADD), testosterone, epitestosterone, 17α- and 17β-nandrolone, and trenbolone in powdered milk for calves. All analytes were extracted, after a common deproteinization and defatting sample pre-treatment, by a unique immunoaffinity column and analysed by liquid chromatography tandem mass spectrometry (LC–MS/MS) in both positive and negative electrospray ionization (ESI) modes. The method was validated according to the criteria of the Commission Decision 2002/657/CE. The analytical limits were from 0.39 to 0.73 ng mL−1 for the decision limit (CCα) and 0.46–0.99 ng mL−1 for detection capability (CCβ). The analysis of 50 samples of milk replacers for calves, always revealed the presence of cortisol and cortisone (average concentrations 2.56 and 1.06 ng mL−1, respectively), frequently testosterone and epitestosterone (1.24 and 0.63 ng mL−1, respectively), occasionally β-nandrolone (0.82 ng mL−1) and prednisolone (0.41 ng mL−1). The other anabolic steroids were never found.