Quantitation of tizoxanide in multiple matrices to support cell culture, animal and human research

•The liquid chromatography mass spectrometry assay is validated for quantification of tizoxanide, the active metabolite of nitazoxanide, in human plasma, mouse plasma and Dulbeccos Modified Eagle Medium containing FBS.•The impact of matrix effects on quantification of tizoxanide in the assay is minimal.•The validated assays linear range is between 15.6 ng/mL-1000 ng/mL.