BACKGROUND: We have previously demonstrated the isoflurane and halothane may be detrimental to in vitro fertilization of mouse oocytes in high concentrations. The aim of this study is to compare the toxic effects of volatile anesthetics on mouse embryos using in vitro growth model of two cell mouse embryos. METHODS: Mouse two-cell embryos exposed to three volatile anesthetics, enflurane(0.5 mM; 1.5 mM), isoflurane(0.26 mM; 0.78 mM) and halothane(0.24 mM; 0.72 mM). Mouse two-cell embryos unexposed to any drugs were included as controls. RESULTS: The percentages of two-cell mouse embryos developed over morula stages on the third day after exposure of high concentrations of isoflurane and halothane decreased significantly compared with controls. The rates of embryos arrested at 2-8 cell stage in these groups were significantly higher than that of controls. There were no significant differences in these rates between enflurane group, isofiurane and halothane group of lower concentrations and controls. The hatching and/or hatched blastocysts development were significantly lower in isoflurane and halothane group than in controls. No significant differences in the hatching rate of blastocyst developed were observed among groups. CONCLUSIONS: Our data show that isoflurane and halothane in high concentrations have harm effects of the in vitro growth of two cell mouse embryos.