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A toxicity evaluation of two Keggin-type heteropolytungstates, K7[Ti2PW10O40]·6H2O and K6H[SiV3W9O40]·3H2O, with different inhibitory potencies toward acetylcholinesterase activity (IC50 values of 1.04 × 10− 6 and 4.80 × 10− 4 mol/L, respectively) was performed. Wistar albino rats were orally treated with single doses (5 and 50 mg/kg) of both investigated compounds. The biochemical parameters of renal (serum urea and creatinine) and liver function (direct and total bilirubin, alanine transaminase, and aspartate aminotransferase) were determined after 24 h and 14 days. A histopathological analysis of liver tissue was carried out 14 days after the polyoxotungstate administration. Both applied doses of the investigated compounds did not induce statistically significant alterations of the renal function markers. However, the polyoxotungstate treatment caused an increase in the activities of serum alanine transaminase and aspartate aminotransferase in a time- and concentration-dependent manner, although statistically significant changes in bilirubin concentrations were not observed. Furthermore, the detected hepatotoxic effect was confirmed by histhopathological analysis that suggested some reversible liver tissue damage two weeks after the treatment, especially in the case of K6H[SiV3W9O40]·3H2O. Accordingly, the toxicity of these two polyoxotungstates with anti-acetylcholinesterase effect cannot be considered as a severe one, but their potential clinical application would require a more complex toxicological study.K-Ti2PW10 and K-SiV3W9in vitro inhibit AChE activity with different potencies.Both K-Ti2PW10 and K-SiV3W9 do not influence rat body mass and food intake.Both K-Ti2PW10 and K-SiV3W9 do not significantly alter biomarkers of renal function.K-Ti2PW10 and K-SiV3W9 significantly increase hepatotoxicity parameters, ALT and AST.K-Ti2PW10 and K-SiV3W9 cause reversible liver tissue damage 2 weeks after application.