A simple and specific analytical method for the quantitative determination of five cucurbitane-type triterpenoids isolated from the fruit of Momordica charantia is developed. The triterpenoids present in the fruits of Momordica charantia are separated with an acetonitrile (0.1% acetic acid)-water (0.1% acetic acid)-methanol (0.1% acetic acid) gradient at a flow rate of 0.5 mL/min. The high-performance liquid chromatography separation was performed on a Phenomenex C18 reversed-phase column. By using an evaporative light scattering detector, the main triterpenoids of Momordica charantia could be detected at levels as low as 10 microg/mL. The method was validated for precision, repeatability, and accuracy. The relative standard deviation was between 0.6-4.4%. The method was sensitive, quick, and accurate for the determination of main triterpenes and saponins in Momordica charantia, and can be used for quality control of Momordica charantia and its related dietary supplements.