The impact of high hydrostatic pressure (HHP) combined with moisture regulators (MR) on the quality and flavor attributes of beef jerky during storage was investigated in this study. During storage, moisture content decreased in all the treatments, and it was significantly higher in MR + HHP samples than in HHP and control samples (P < 0.05). At 0 and 15 days, the water activity was approximately 0.7, and at 60 days, it decreased to 0.652 for control samples and 0.678 for MR + HHP samples. During storage, the MR + HHP samples had lower shear force and thiobarbituric acid reactive substances (TBARs) value than the control and HHP samples (P < 0.05). MR + HHP increased the formation of volatile compounds derived from spices while inhibiting the formation of volatile compounds derived from lipid oxidation. The sensory scores of MR + HHP samples for tenderness, color, taste, and flavor were significantly higher than that of the HHP and control samples (P < 0.05). This study revealed that HHP and MR treatment reduced the quality deterioration of beef jerky during storage.