Direct determination of nosiheptide residue in animal tissues by liquid chromatography-tandem mass spectrometry

Because only very weak signals of fragment ions of nosiheptide can be obtained, nosiheptide is usually detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) via the determination of its hydrolyzed degradation product named HMIA in previous studies. The indirect method suffers from several problems, such as complicated samplepreparation, unavailable commercial HMIA, and the risk of the false-positive result by HMIA. However, we found that nosiheptide could produce several significant fragment ions under high collision energy (CE). Therefore, we developed a method for the direct determination of nosiheptide by LC-MS/MS in animal tissues. The sample was extracted with ACN, then degreased with n-hexane, and purified by an HLB solid-phase extraction (SPE) cartridge. After being filtered through the PTFE filter, it was analyzed by LC-MS/MS in selected reaction monitoring (SRM) mode. The influencing factors, such as mobile phase, SPE cartridge, filter material, and matrix effect, were investigated. Nosiheptide showed a good linear relationship (R² ≥ 0.999) within the concentration range from 0.3 μg/L to 20 μg/L under optimized conditions. The limit of detection (LOD) was 0.3 μg/kg, while the limit of quantification (LOQ) was 1.0 μg/kg in chicken, bovine muscle, swine muscle, and swine liver. The average recoveries at spiked levels of 1.0, 2.0, and 10 μg/kg ranged from 83% to 101%, with the relative standard deviations (RSDs) <12%. Compared with the methods previously reported, our newly developed method was more simple, convenient, and sensitive. Moreover, it was successfully applied for the determination of nosiheptide residue in medicated chicken samples.