Effects of combinations of methanol and formic acid on rat embryos in culture.

Acute human exposure to methanol (MeOH) results in elevated serum concentrations of both MeOH and formic acid. In order to better assess the risk of adverse developmental effects of MeOH exposure in humans, the effects of the combination of formate and MeOH, in addition to the individual toxicity profiles for MeOH and formate, need to be established. Gestational day 9 rat embryos were exposed to various concentrations of MeOH and formate in whole embryo culture (WEC) for 48 hr and the degree of embryotoxicity was evaluated using developmental score (DEVSC) as the parameter of comparison across exposure combinations. After establishing embryo toxicity of the individual compounds in previous studies, concentrations of MeOH and formate were chosen which would produce similar DEVSCs, and isoboles were plotted joining the equivalently toxic doses. These mixtures would be expected to have similar toxicity to the MeOH or formate concentrations according to a dose-addition model. The responses of embryos to the selected concentrations along each isobole were measured and tested for linearity to determine the nature of any interaction between the two agents. The concentrations of MeOH and formate used separately and in combination ranged from 0 to 8.75 mg/ml MeOH and 0 to 1.51 mg/ml formate. Increasing concentrations of either MeOH or formate resulted in significant decreases in DEVSC. Exposure to combinations of MeOH and formate had less effect on DEVSC than would be expected based on simple toxicity additivity. This observation was also true for embryonic crown-rump length, head length, and somite number. These results suggest that the embryotoxicity observed following low level exposure to MeOH is mechanistically different from that observed following exposure to formate.