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Article Title

Confirmatory method for nine sulfonamides in miniature bovine muscle samples using HPLC/MS/MS without using SPE

Abstract

A new confirmatory method for the analysis of nine sulfonamides (sulfamethazine, sulfamethizole, sulfamethoxypyridazine, sulfamethoxazole, sulfapyridine, sulfachlorpyridazine, sulfamethazine, sulfamethoxine and sufaquinoxaline in small bovine muscle samples (0.1 g) is presented. The method, based on HPLC/MS/MS, identifies and quantifies the sulfonamides in bovine muscle at concentrations below the maximum residue limits (100 ng/g), established by the European Commission and by the Unites States Food and Drug Administration. The extraction protocol is rapid and easy, and does not require the use of solid phase extraction (SPE) cartridges. Validation was conducted according to the Commission Decision 2002/657/EC with fortified bovine samples and employing sulfadoxine-d3 as internal standard. The linearity of the method was observed at the concentration range between 25 and 250 ng/g. Intra-day and inter-day recoveries obtained in the validation of the method were above 88 % for all sulfonamides. Applicability of the method was investigated in bovine muscle samples belonging to a regional control program and results were compared with those obtained with an HPLC/DAD method accredited by ENAC (National Spanish Entity for Accreditation).

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