A CCl4/CHCl3 interaction study in isolated hepatocytes: selection of a vehicle

Emulphor, ethanol, and dimethyl sulfoxide (DMSO) were evaluated as vehicles in studying the toxicity of CCl4 and CHCl3 in isolated hepatocytes. The appropriateness of the vehicle was determined by evaluating the following parameters: solubility of CCl4 and CHCl3 in the vehicle, cell injury (intracellular K+), cell death (LDH leakage), and lack of interaction (protection or enhanced toxicity) with CCl4 and CHCl3. The relative toxicity of the vehicles according to maximum no effect levels (v/v) was: emulphor (0.125%) greater than ethanol (1.0%) greater than DMSO (5.0%). Emulphor at toxic levels was inadequate to dissolve enough CCl4 to evaluate in this system. Ethanol (5.0, 2.5, 1.0, 0.5%) was more toxic than DMSO and interacted with both CCl4 and CHCl3 to enhance toxicity. DMSO (15.0, 5.0, 2.5%) did not significantly alter the toxicity of CCl4 and CHCl3; no interaction. These data suggest that DMSO should be the vehicle for evaluating the toxicity of CCl4 and CHCl3 and their mechanisms of action in the isolated hepatocyte.