This study employed the response surface methodology to optimize the extraction conditions for recovering vitamins D2 and K1 from green leafy vegetables using ultrasonication-assisted extraction. The vitamin content was determined using an Accucore C18 column and a UPLC-Q-ToF/MS method. An RSM-I-Optimal design was used for designing the experiment to find the best combination of solvent level (mL), sonication time (min), sonication frequency (kHz), and temperature (°C). The experimental data from a 25-sample set were fitted to a second-order polynomial equation using multiple regression analysis. The extraction models had R2 values of 0.895 and 0.896, respectively, and the probability values (p < 0.0001) indicated that the regression model was highly significant. The optimal extraction conditions were: solvent level of 65 mL, sonication time of 45 min, sonication frequency of 70 kHz, and temperature of 45 °C. Under these conditions, the predicted recovery (%) values for vitamins D2 and K1 were 90.7% and 90.4%, respectively. This study has the potential to use the reported extraction method for routine quantification of vitamins D2 and K1 in the laboratory using UPLC-Q-ToF/MS.