Following the ban of four nitrofurans in the mid-90s (furazolidone, furaltadone, nitrofurantoine, nitrofurazone), the nifursol, a veterinary drug from the nitrofuran class of antibacterials which has been used prophylactically as feed additive for treating turkeys against histomoniasis (blackhead disease) was also declared in Annex IV of the European Union Directive no. 90/2377/EC in 2002 according to the Regulation no. 1756/2002/EC. As for the four other nitrofurans, nifursol disappears from tissues within a few days after treatment of food-producing animals. But toxic metabolites are still present for longer periods (several weeks or even months). The major metabolite that can readily be monitored in the tissues following nifursol abuse is the 3,5-dinitro-salicylic acid hydrazine (DNSAH). This article displays some improvements and the revalidation of the analytical method by liquid chromatography coupled to electrospray tandem mass spectrometry (LC-esiMS/MS) already in use in our laboratory for monitoring nitrofuran metabolites but also including the nifursol metabolite at the confirmatory minimum required performance level (MRPL) of 1 microg kg(-1). The validation is applied both to artificially and to naturally incurred turkey muscle.