Sea nettle (Chrysaora quinquecirrha) venom (CQV) is known to be toxic to the cardiac, respiratory, renal and hepatic systems in animal models. However, the mechanism of toxicity of CQV on hepatocytes is unknown. We utilized isolated rat hepatocytes in culture and measured percentage total lactate dehydrogenase (LDH) release after direct exposure to CQV. Toxicity of CQV in this system produced a linear multiple-dose response curve as well as a linear single-dose kinetics curve. Neither extracellular calcium concentration nor intracellular calcium chelation had a statistically significant effect on the toxicity of CQV in our hepatocyte model. From these results it appears that CQV does not form large membrane channels similar to complement, nor does calcium appear to play a major role in the mechanism of toxicity in hepatocytes. The isolated rat hepatocyye culture and measure of LDH release provided a relatively simple and reproducible model for examining toxicity of CQV.