Optimization of injected gallium-68 (68Ga) activity for 68Ga-prostate-specific membrane antigen positron emission tomography/computed tomography (68Ga-PSMA PET/CT) studies is relevant for image quality, radiation protection, and from an economic point of view. However, no clear guidelines are available for 68Ga-PSMA studies. Therefore, a phantom study is performed to determine the highest coefficient of variation (COV) acceptable for reliable image interpretation and quantification. To evaluate image interpretation, the relationship of COV and contrast-to-noise ratio (CNR) was studied. The CNR should remain larger than five, according to the Rose criterion. To evaluate image quantification, the effect of COV on the percentage difference (PD) between quantification results of two studies was analyzed. Comparison was done by calculating the PD of the SUVmax. The maximum allowable PDSUVmax was set at 20%. The highest COV at which both criteria are still met is defined as COVmax. Of the NEMA Image Quality phantom, a 20 min/bed (2 bed positions) scan was acquired in list-mode PET (Philips Gemini TF PET/CT). The spheres to background activity ratio was approximately 9:1. To obtain images with different COV, lower activity was mimicked by reconstructions with acquisition times of 10 min/bed to 5 s/bed. Pairs of images were obtained by reconstruction of two non-overlapping parts of list-mode data. For the 10-mm diameter sphere, a COV of 25% still meets the criteria of CNRSUVmean ≥ 5 and PDSUVmax ≤ 20%. This phantom scan was acquired with an acquisition time of 116 s and a background activity concentration of 0.71 MBq/kg. Translation to a clinical protocol results in a clinical activity regimen of 3.5 MBq/kg min at injection. To verify this activity regimen, 15 patients (6 MBq/kg min) with a total of 22 lesions are included. Additional reconstructions were made to mimic the proposed activity regimen. Based on the CNRSUVmax, no lesions were missed with this proposed activity regimen. For our institution, a clinical activity regimen of 3.5 MBq/kg min at injection is acceptable, which indicates that activity can be reduced by almost 50% compared with the current code of practice. Our proposed method could be used to obtain an objective activity regimen for other PET/CT systems and tracers.