Development and validation of a multi-residue method for determination of 18 β-agonists in bovine urine by UPLC-MS/MS

Ultra performance liquid chromatography (UPLC) hyphenated to tandem mass spectrometry (MS/MS) was used for the development of an analytical method capable of simultaneous identification and quantification of 18 β-agonist compounds namely, brombuterol, chlorbrombuterol, cimaterol, cimbuterol, clenbuterol, clencyclohexerol, clenisopenterol, clenpenterol, clenproperol, hydroxymetylclenbuterol, isoxsuprine, mabuterol, mapenterol (clenbuterol-like compounds), ractopamine, ritodrine, salbutamol, salmeterol (salbutamol-like compounds) and zilpaterol in bovine urine. In compliance with the Commission Decision 2002/657/EC (CD 2002/657/EC), the method was validated applying a matrix-comprehensive in-house validation approach based on a fractional factorial design. Six experimental factors varied on two levels were selected for this purpose. The relevant validation parameters such as decision limit CCα (range, 0.24-0.51 μg L− 1) and detection capability CCβ (range, 0.27-0.71 μg L− 1), within-laboratory reproducibility (< 20%) as well as recovery (range, 92-109%) were in agreement with the performance criteria set in the CD 2002/657/EC. Overall, the proposed method enabled both screening and confirmatory detection of the β-agonist compounds in the framework of official monitoring plans.