Determination of dinitolmide and its metabolite 3-ANOT in chicken tissues via ASE-SPE-GC–MS/MS

Abstract A novel confirmatory analysis method to simultaneously extract, clean-up and detect dinitolmide and its metabolite (3-amino-2-methyl-5-nitrobenzamide, 3-ANOT) in chicken tissues using accelerated solvent extraction (ASE) with a neutral alumina solid-phase extraction (SPE) column coupled with gas chromatography-tandem mass spectrometry (ASE-SPE-GC–MS/MS) was established. GC–MS/MS was performed using a VF-5ht capillary column with helium as the carrier gas, temperature programming, and an inlet temperature of 340 °C. The experimental method was carried out in the electron ionization mode; the full scan mode was used for qualitative analysis, and selective reaction monitoring, combined with an external standard method was used for quantitation. Under the optimized conditions, at the addition levels of the limits of quantitation (LOQs), 0.5 MRL (maximum residue limit), 1 MRL and 2 MRL, the average recoveries of dinitolmide and 3-ANOT in chicken tissues were 81.96–94.31%, and the relative standard deviations (RSDs) were 1.72–5.37%. The limits of detection (LODs) (S/N ≥ 3) of dinitolmide and 3-ANOT in chicken tissues were 0.8–2.5 μg/kg, and the LOQs (S/N ≥ 10) were 2.7–8.0 μg/kg. The method validation parameters met the Chinese Ministry of Agriculture, European Union and United States Food and Drug Administration veterinary drug residue detection requirements.