This study was targeted to investigate the effect of the combination of aqueous ozone (AO) and sodium citrate (SC) pretreatment on quality of vacuum packaged beef stored at chilling temperature. The steers were randomly divided into five treatment groups, containing the group C (steers without any treatment), DW (steers pretreated with distilled water), AO, SC and AO+SC. During processing, the parameters included total Viable Count (TVC), total Volatile Basic Nitrogen (TVB-N), pH, meat color, metmyoglobin (Met-Mb), thiobarbituric acid reactive substances (TBArs), Warner-Bratzler shear force (WBSF), and purge loss were measured and compared. Firstly, the TVC in beef steaks of group AO+SC was significantly lower than group AO and SC during the whole storage (P < .05). In addition, the TVB-N values in AO+ SC group were significantly decreased than AO group at the end of storage time. Furthermore, compared with the group only treated by AO, the group AO+SC displayed higher L* (lightness) value, lower a* (redness) value and lower Met-Mb. The values of WBSF and purge loss of group AO+SC was lower than group AO, as well as TBArs. The treatment methods and storage time had significant interactive effects on TVC, TVB-N, pH, L*, Met-Mb, and TBArs (P < .05). The combination treatment of AO and SC showed better effect on inhibiting the microbial growth and decreased the deteriorated effect of AO on beef quality including TBArs, WBSF, and purge loss.