A rapid high performance liquid chromatographic method for the determination of etoposide (VP16) in human serum has been developed. The samples were added teniposide (VM26) as an internal standard, extracted with ethyl acetate and chromatographed on a reversed phase YWG-C18 column with UV detection at 254 nm. The mobile phase was CH3OH:H2O = 55:45 (V/V). The calibration curve was Y = 0.0160 + 0.034X, r = 0.9991. It was reticlinear in the range between 0.5-50 mg/L. The detection limit was 0.1 mg/L. The average recovery was 95.0% +/- 0.2%. Intra-day and inter-day RSD were 4.16% and 4.76% respectively. From the results it can be concluded that this method is reliable. This method has been used to determine VP16 concentration in patient serum. The data obtained showed that the sensitivity and selectivity of this method were adequate for drug monitoring in clinical research.