The quality changes of RJ at different storage conditions are evaluated from various perspectives, including non-target volatile compound profiling, multi-index (physicochemical parameters and flavor characteristics), and multi-technology (HS-SPME-GC/MS, electronic nose, and electronic tongue) integration. Display omitted •A total of 37 volatile compounds in royal jelly are identified using HS-SPME-GC–MS.•Seven volatile compounds contribute largely to the overall flavor of royal jelly.•Electronic nose/tongue exhibit useful value in the quality control of royal jelly.•Combination of multiple methods enhance the evaluation of royal jelly quality. Royal jelly (RJ) is known for its unique flavor and nutritional value. However, the flavor changes of RJ during storage remain unclear. In this work, the flavor profiles of RJ during storage were evaluated by using headspace solid-phase micro-extraction gas chromatography-mass spectrometry (HS-SPME-GC/MS) combined with both electronic nose and electronic tongue analyses. Results revealed that the moisture, water-soluble protein, and whiteness were changed significantly at 25 °C. The holistic variation of RJ flavor exhibited evident distinction based on principal component analysis with electronic nose and electronic tongue. Among the total of 37 volatile compounds identified in RJ, the octanoic acid showed the highest contents of 47.61 % at 25 °C in 21 d. Seven volatile compounds, i.e., 2(3H)-furanone,5-butyldihydro-, 2-heptanone, trans-β-ocimene, 2-nonen-4-one, 2-nonanone, methyl benzoate, and 2-octenoic acid (E), contributed largely to the typical overall flavor of RJ. This work provides an improved understanding of the flavor change of RJ during storage.