Food sources foraged by bees are a key external factor influencing the chemical composition of royal jelly. However, existing research has primarily focused on its nutritional value and biological activity, and studies on the chemical determinants underlying the taste of royal jelly remain limited. Thus, to objectively characterize the taste profiles and taste-related compounds of royal jelly produced from different food sources, the authors employed standard methods to measure acidity, sugars, and fatty acids to determine the composition and contents of taste-related compounds. Taste differences were analyzed using electronic tongue technology. The results revealed that royal jelly produced by bees fed natural bee pollen mixed with honey (PH group) exhibited the most pronounced sweetness and umami characteristics, whereas royal jelly from bees fed natural bee pollen mixed with sugar water (PS group) had strong sourness and bitterness. Furthermore, royal jelly from bees fed substitute pollen mixed with honey (TH group) was characterized by a distinct umami taste and relatively low sourness. Sugar analysis revealed that the PS group produced royal jelly with significantly lower total sugar, glucose, and fructose contents (10.09, 4.36, and 4.05 g/100 g, respectively) than the PH and TH groups (P<0.05). Conversely, sucrose and erlose concentrations in the PS group (1.69 and 0.48 g/100 g) were significantly higher than those in the other two groups. Fatty acid analysis revealed that royal jelly from the PH group contained the highest total fatty acid content (4.46 wt.%), which was significantly higher than that of the PS and TH groups (P<0.05). The PH group also exhibited the highest levels of C16:0 and C16:1 fatty acids and the lowest levels of C18:1 (cis-n9). In contrast, royal jelly from the TH group contained significantly higher levels of C21:0 and significantly lower levels of C18:3n3 than that from the PH and PS groups, respectively (P<0.05). In addition, the concentration of 10-hydroxy-2-decenoic acid (10-HDA) (1.56 wt.%) in jelly from the PS group was significantly lower than that observed in the PH and TH groups (P<0.05); however, C24:0 was not detected at all. Furthermore, C18:1 (cis-n9) and C21:0 were found to show a significant negative correlation with the sourness, sweetness, and saltiness of jelly, whereas 10-HDA and C16:1 showed a significant positive correlation with sourness. However, fatty acid composition appeared to have only a minimal influence on bitterness and umami taste attributes. In conclusion, the sources of food available to bees influence the taste of royal jelly by altering the compositions and concentrations of taste-related compounds. The findings of this study will provide a theoretical basis for evaluating the taste, identifying the origin, and improving the sensory qualities of royal jelly.