A galactose‐ and N‐acetyl‐D‐galactosamine‐specific lectin (Viscum album L. var. coloratum agglutinin, VCA), which is known for its anticancer activity, was isolated from mistletoe. In this study, we investigated the antimutagenic potentials of VCA by using the pre‐incubation method of the Ames test (Salmonella typhimurium TA98 and TA100) in the presence or absence of S9 mixture. Viscum album L. var. coloratum agglutinin was assessed for its antimutagenic properties against the mutagens 2‐aminoanthracene (2AA) and furylfuramide (AF‐2) for strain TA98, and sodium azide (NaN3) and 2‐aminoanthracene (2AA) for strain TA100. The concentrations used for this test compound were 100, 200 and 400 µg per plate. Viscum album L. var. coloratum agglutinin showed moderate, but not negligible, protective effects regarding the antimutagenic properties against the direct‐acting mutagens NaN3 and AF‐2. Furthermore, VCA was more effective in preventing the mutagenicity of the indirect‐acting mutagen 2‐AA (in the presence of S9) when tested with both TA98 and TA100. In conclusion, this report has shown broad ranging antimutagenic effects of VCA to numerous mutagens in TA98 and TA100 Salmonella typhimurium strains. Although the data presented here cannot be applied in vivo, they can support other antimutagenic and anticarcinogenic findings for VCA. Copyright © 2011 John Wiley & Sons, Ltd.