Beef potentiator is an important flavor enhancer in food industry, while it is prone to generating insufficient taste compounds and off-odor compounds during enzymatic hydrolysis of beef, resulting in poor flavor of beef potentiator. It has been extensively reported that sonication is capable of improving food flavor. However, the effect of sonication on the flavor of enzymatically hydrolyzed beef liquid (EHBL) was scarcely reported. Herein, we investigated the effect of sonication on the flavor of EHBL using quantitative descriptive analysis (QDA), physicochemical analysis and SPME-GC-olfactometry/MS. QDA showed that sonication had a significant effect on taste improvement and off-odor removal of EHBL. Compared with the control, sonication (40 kHz, 80 W/L) increased the contents of total nitrogen, formaldehyde nitrogen, total sugar, reducing sugar, free amino acids (FAAs) and hydrolysis degree of EHBL by 19.25%, 19.80%, 11.83%, 9.52%, 14.37% and 20.45%. Notably, sonication markedly enhanced the contents of sweet FAAs, umami FAAs and bitter FAAs of EHBL by 19.66%, 14.04% and 9.18%, respectively, which contributed to the taste improvement of EHBL. SPME-GC-olfactometry/MS analysis showed that aldehydes and alcohols were the main contributors to aroma compounds of EHBL, sonication significantly increased the contents of key aroma compounds and 115.88% of alcohols content in EHBL. Notably, sonication decreased contents of the fishy odorants, hexanoic acid and nonanal, markedly by 35.29% and 26.03%, which was responsible for the aroma improvement of EHBL. Therefore, sonication could become a new potential tool to improve the flavor of EHBL.