Clostridium difficile is an anaerobic bacillus responsible for intestinal disease associated with previous antibiotic treatment. The transmission of the disease is associated to hospital environments contamination and possible association with the consumption of contaminated foods. The objective of the study was to evaluate the sporicidal activity of sanitizers on spores of C. difficile, according to the European Union method prEN14347. The sanitizers used were: 1% sodium hypochlorite, pH 11.8; 0.03% peracetic acid; 0.45% Alkyl amido propyl dimethyl benzyl ammonium chloride and 0.03% polyhexamethylene biguanide hydrochloride; the strains tested were VPI 10463 (clinical origin) and LHCD 809 (food origin). The sporicidal effect of sanitizer was evaluated using the inactivation factor (IF). Peracetic acid promoted a reduction in spore counts (IF 2.69 ± 0.20 and 2.39 ± 0.11 log CFU/mL for C. difficile VPI 10463 and LHCD 809, respectively) after 30 minutes but did not show sporicidal effect (IF < 4.0 log CFU/mL). Only 1% sodium hypochlorite after 30 minutes of contact showed a sporicidal effect on strain C. difficile VPI 10463, with IF 4.20±0.20 log CFU/mL and reduction of spore counts of strain LHCD 809 (IF 3.24 ± 0.21 log UFC/mL) after 30 minutes of contact. Biguanide and quaternary ammonium had no effect on C. difficile spores. The results, have evidenced the inefficacy of some commercial sanitizers to eliminate C. difficile spores.