One of the main practices followed by beekeepers during the production of royal jelly (RJ) is the artificial sugar feeding. In this study, the effect of carbohydrate supplementary feeding on the composition of the three main sugars (fructose, glucose, sucrose) and on the final quantity of the product was explored using one-way ANOVA and non-parametric tests. Also, the correlations among the parameters were examined. The average yield per colony for non-supplemented colonies (Group A) was 12.8 g, while the average content of fructose, glucose and sucrose was 4.32%, 3.78%, and 0.04%, respectively. For the colonies fed at the grafting day one time (Group B), these values were 12.76 g, 3.11%, 3.19% and3.71%, and for the colonies fed from the insertion until the collection day (Group C), 12.81 g, 3.05%, 3.12% and 3.54% respectively. It should also be noted that the sucrose content in all samples from supplemented colonies was found greater than 1.97%. The statistical tests highlighted the impact of artificial feeding on fructose and glucose contents, while the produced quantity remained uninfluenced. Finally, the Spear man (rho) coefficient test showed statistically significantly negative correlation between the monosaccharides (fructose, glucose) and sucrose.