Smokeless tobacco products, and in particular snus, are being increasingly considered as Potential Reduced Exposure Products, and in some quarters as smoking cessation products. Although epidemiological evidence from Sweden suggests that snus use is substantially less hazardous than cigarette smoking, snus is not considered as harmless. The objective of this work described was to establish an appropriate extraction methodology for Snus such that robust safety assessment of snus products could be conducted in the Ames, Mouse Lymphoma, in vitro Micronucleus and Neutral Red Uptake assays. There is no standard method for applying snus to in vitro test systems, but most workers use snus extractions to provide treatment solutions compatible with the assay systems. We have applied certain modifications to the ISO 10993-12 guidelines (for the biological evaluation of insoluble medical devices) in order to prepare snus extracts which allow robust safety assessment in various in vitro genotoxicity and cytotoxicity assays. Snus samples were extracted for 24 hours at 37°C in both water and dimethyl sulphoxide (DMSO), at varying concentrations up to 500 mg (equivalent) per mL. Nicotine content was measured as 82-86% and 73% for aqueous Snus extracts at 200 and 500 mg (equivalent) per mL respectively, indicating relatively little loss in relative nicotine recovery over this concentration range. Extraction at a higher concentration was considered impractical due to excessive absorption of the extraction vehicle. In vitro genotoxicity assays performed using snus extracts at this highest practical concentration exceed maximum exposure concentrations required by the standard regulatory guidelines applicable for these assays. In some cases treatments resulted in biological dose limiting effects, further demonstrating the rigour of these safety assessments.