The chlorinated acetates monochloroacetate (MCA), dichloroacetate (DCA), and trichloroacetate (TCA) are generated in water disinfection processes and are formed during metabolic detoxification of industrial solvents such as trichloroethylene. In order to test for consequences of an exposure of brain cells to the different chlorinated acetates, glutathione levels and lactate production of primary astrocyte cultures were investigated as indicators for the potential of chlorinated acetates to disturb cellular detoxification processes and glucose metabolism, respectively. Application of MCA to cultured astrocytes caused a time and concentration dependent deprivation of cellular glutathione, inactivation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity, and loss in cell viability with halfmaximal effects observed for MCA concentrations between 0.3 and 3 mM. In contrast, the presence of acetate, DCA, or TCA in a concentration of 10 mM did not compromise cell viability nor affect cellular glutathione content or GAPDH activity. However, the presence of DCA and TCA significantly lowered the rate of cellular lactate production in viable astrocytes. These data demonstrate that the extent of chlorination strongly determines the potential of chlorinated acetates to disturb glutathione and/or glucose metabolism of astrocytes.