Abstract: The present study investigated the effects of ultrasonic waves assisted vacuum tumbling on eating quality of beef meatballs. Taking the traditional treatment group (CON) as the blank control, the beef surimi was treated by vacuum tumbling (VT), ultrasonic waves (UW) and ultrasonic waves assisted vacuum tumbling (VT+UW) respectively, and then uniformly made into beef meatballs. The cooking loss, texture characteristics, microstructure and volatile flavor of meatballs treated with different processing methods were measured. Results showed that the cooking loss of beef meatballs was significantly decreased (P<0.05) by the three treatments. VT+UW treatment could significantly improve the hardness, gumminess and chewiness of meatballs, and reduce the adhesion (P<0.05). The result of paraffin section demonstrated that VT+UW treatment could promote the formation of a denser inner gel network structure. The total amount of volatile flavor substances in beef meatballs identified by gas chromatograph-mass spectrometry (GC-MS) was up to 16.16 mg/kg after 10 min of ultrasonic waves assisted vacuum tumbling treatment (VT+UW1). Moreover, the contents of hydrocarbons in VT+UW treatment group increased, while the contents of aldehydes and alcohols decreased. The relative odor activity value (ROAV) showed that nonanal, (Z)-3,7-dimethyl-2,6-octadienal, (+)-limonene and O-isopropylbenzene were the characteristic flavor substances of beef meatballs, and reached a relatively higher content in VT+UW group. It was further confirmed that VT+UW treatment was more beneficial to improve the flavor of beef meatballs by enriching the characteristic flavor compounds. The results indicated that suitable VT+UW treatment could not only reduce the cooking loss of beef meatballs, but also improve the texture and flavor characteristics of beef meatballs.