Traditional sturgeon slaughter practices, involving a two-step process of percussive stunning followed by exsanguination, often fail to deliver humane outcomes efficiently due to the robust and cartilaginous anatomy of these large fish, which typically necessitates multiple strikes to achieve insensibility. This inefficiency not only complicates the slaughter process but also raises significant welfare concerns, highlighting the urgent need for improved methods. In response, this study evaluates the efficacy of a non-penetrative captive bolt (NPCB) gun as an alternative method aimed at enhancing animal welfare during the slaughter of farmed white sturgeon (Acipenser transmontanus). Utilizing a Jarvis HPS-1 NPCB, the research applied concussive strikes at various pressures (120, 135, and 145 PSI for juveniles, and 175, 200, and 225 PSI for adults) prior to exsanguination and immersion in an ice bath. Subsequent histological examinations of cartilage sections from the brain area were conducted to assess the extent of physical damage and to correlate strike pressures with intracranial hemorrhage. The results indicated a significant relationship between increased pressure and the occurrence of meningeal-to-cerebral hemorrhage in juveniles, with no significant differences observed in adults. Recovery monitoring revealed that 100% of juveniles and 90% of adults did not recover post-stunning with exsanguination, validating the NPCB’s potential to maintain insensibility effectively. However, the absence of immediate brain death in some cases suggests that while NPCB stunning significantly improves upon traditional methods by reducing the need for multiple manual strikes, further research is essential to refine this technique. Optimizing pressure settings is crucial to consistently achieve immediate brain death with a single strike, aligning the slaughter process with the highest standards of animal welfare and addressing the unique challenges posed by the anatomical characteristics of sturgeon.