Method development and validation for the simultaneous determination of 21 antiviral drugs by ultra-high performance liquid chromatography-tandem mass spectrometry in chicken muscle and liver

The recent unauthorization of antiviral drugs in food-producing animals according to Commission Delegated Regulation (EU) 2022/1644 have increased the need for food control laboratories to develop analytical methods and perform official controls. In this work, a simple and fast analytical methodology was developed for the simultaneous determination of 21 antiviral drugs in chicken muscle and liver by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Chromatographic separation was achieved by an HILIC BEH amide column; followed by detection with a electrospray ionization source in positive and negative modes. Based on extraction efficiencies, critical parameters affecting sample treatment were optimized including the evaporation and clean up steps to extract the largest number of antiviral drugs and reduce interferences. The method was validated according to Commission Implementing Regulation (EU) 2021/808 in chicken muscle and liver. Most compounds showed a linearity of R²>0.9800, while decision limits were between 0.18 and 7.05 μg kg⁻¹ and 0.19 and 36 μg kg⁻¹ for chicken muscle and liver, respectively. Trueness and within-lab reproducibility were determined at three levels (n = 7) and the results showed values ranging from 81 to 133 % and 4.2–57 % for chicken muscle, and 71–136 % and 4.6–106 % for chicken liver, respectively. The applicability of the developed method was demonstrated by the analysis real samples. 20 samples from the National Residue Control Plan in the Netherlands were analyzed and although none of targeted compounds were detected it is important to continue the analysis of larger set of samples to address any possible food safety risks.