Abstract: A coupled UPLC-MS/MS method has been developed and validated for the simultaneous quantitation of food sweeteners stevioside (STV) and aspartame (ASP). Good chromato-graphic separation was achieved on a Hypersil gold 50×2.1 mm (1.9 μm) column, using a gradient flow of 10 mM ammonium acetate, pH = 2.9 adjusted with formic acid, and 10 mM ammonium acetate in acetonitrile:water (95:5, v:v) as the mobile phase. The eluate was introduced to ESI-Mass spectrometer and scanned using multiple reaction monitoring (MRM). The method was robust, reproducible and easy to use and was validated according to ICH guidelines for the accuracy and precision giving acceptable ranges. The utilization of multiple reaction monitoring improved the selectivity of detection. Method was linear in the ranges of 2-250 ng/mL for STV and ASP. Application of this method on laboratory mixtures of the selected sweeteners was successful. The using of mass spectrometry make the method selective and measurement did not affect the presence of impurities, additive and other ingredients of detection due to the simplicity and sensitivity of this method allows the utilization of method in quality control of STV and ASP.