Its (MRLs) for veterinary drugs and their metabolites in animal-origin foods. The MRL for LMS in poultry muscle is ten /kg in the European Union [7] along with the Usa [8], plus the MRL for MBZ and its two metabolites, 5-hydroxymebendazole (HMBZ) and 2-amino-5benzoylbenzimidazole (AMBZ), is 60 /kg in South Korea [9]. However, South Korea has no regulations on the MRL of LMS in poultry tissues. The European Union and also the Usa usually do not have regulations around the MRLs of MBZ and its two metabolites in poultry tissues, although the European Union has corresponding regulations for sheep and horses. However, to enhance financial advantages, some breeders fail to stick to the prescribed medication regimen and the withdrawal period through poultry development, resulting in residual drug levels in food exceeding the MRL. Moreover, excessive LMS enrichment within the human physique may cause severe damage, like cutaneous necrotizing vasculitis, granulocyte hypoxia, or effects around the nervous system [10]. MBZ, HMBZ and AMBZ have embryotoxic and teratogenic properties due to inhibition of tubulin and mitosis. Veterinary drug residues are an essential international meals safety concern, and to monitor pharmaceutical residues, specifically in poultry foods, there is a need to create a universal and fast analytical approach that sensitively and accurately detects the volume of veterinary drug residue by straightforward sample preparation. At present, the primary Calphostin C custom synthesis detection procedures for LMS and MBZ are immunoassays, gas chromatography (GC) and liquid chromatography (LC). LC solutions primarily incorporate highperformance liquid chromatography (HPLC), ultra-performance liquid chromatography (UPLC), and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Guo et al. developed a colloidal gold immunochromatographic assay based on universal monoclonal antibodies for the simultaneous detection of benzimidazole drug residues in milk samples [11]. Despite the fact that some studies have used GC for the evaluation of LMS [12] and MBZ and its two metabolites [13], GC isn’t as extensively utilized as LC or LC-MS/MS as a result of its standard properties and low volatility of these drugs. Fluorescence detection is best for fluorescence sensitivity and selectivity, but LMS and MBZ don’t exhibit fluorescence and as a result have to be derivatized prior to evaluation. Ultraviolet detection has the same applicability as fluorescence detection, and hence, LC detection of LMS and MBZ and their metabolites in animal-derived meals has mainly been performed with ultraviolet detection [14] and diode-array detection [15,16]. Mass spectrometry has the benefits of higher recovery, high selectivity and superior repeatability, so it can give correct relative molecular masses, comprehensive fragment structural information, higher qualitative stability, and higher detection efficiency for veterinary drug residues in animal foods. In recent years, there have been an increasing variety of research on HPLC-MS/MS detection of LMS or MBZ and its metabolite residues in animal-derived foods [170], but simultaneous detection solutions for these drugs are hardly ever reported, plus the main matrices happen to be aquatic merchandise [21], beef [22], pork [23] and milk [24]. Related study on other poultry muscles has not been reported. As a result, we created an HPLC-MS/MS process for the simultaneous determination of LMS, MBZ, HMBZ, and AMBZ residues Zabofloxacin Purity & Documentation inside the muscle of poultry (chicken, duck and goose). The effects of various extractants and solid-phase extraction (SPE) cartridg.