Ethyl pyruvate (EP) is a simple aliphatic ester derived from pyruvic acid which is an endogenous metabolite. Although various studies have investigated the antioxidant and anti-inflammatory properties of EP, there has been only limited research into the cytotoxic effect of EP on cancer cells. The aim of this study was to determine the cytotoxic effects of EP on cells representing common cancer types. EP was purchased commercially and intermediate stock solutions were prepared with phosphate buffer saline. The cytotoxic effect of EP on human melanoma (VMM917), cervix (HeLa), breast (MCF-7), lung (A549), liver (HepG2), colon (WiDr) cancer and normal fibroblast (BJ) cells was determined using the MTT assay. Cisplatin was used as a positive control in cytotoxicity experiments. The results showed that EP exhibits selective cytotoxic effect on VMM917 (10.1-fold) and HeLa (3.04-fold) cells compared to BJ cells. This study shows for the first time that EP has a highly selective cytotoxic effect, especially on melanoma and cervix cancer cells. The mechanism of this effect needs to be elucidated by more extensive studies.