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Highlights • The present work involves a study to develop a metabolite-incurred reference material. • Water content and mass fraction of nitrofuran metabolites in chicken were evaluated. • An irradiation step was optimized to reduce the initial microbiological contamination. • The method can be applied to produce a homogeneous and stable reference material. • The uncertainty contributions reached fit for purpose levels. Abstract Certified reference materials (CRM) provide measurements of quality and metrological traceability relative to the International System of Units (SI). The results of a feasibility study for the development of an incurred CRM for the mass fraction of nitrofuran metabolites in chicken muscles are presented in this work. A small-scale batch of material was produced from freeze-dried chicken breast muscles containing residues of 3-amino-2-oxazolidinone (AOZ) and 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) metabolites after the controlled in vivo administration of furazolidone and furaltadone antibiotics. A small-scale batch from a blank matrix, without the metabolites, was also produced. The granulometry, stability of residual moisture content, effect of the treatment by irradiation at 3 kGy, 5 kGy and 7 kGy, and degree of inhomogeneity of the property values and residual moisture were evaluated. All statistical approaches for homogeneity and stability assessments were performed based on the principles of ISO Guide 35:2006. [ABSTRACT FROM AUTHOR]