The American Veterinary Medical Association stipulates that appropriate slaughter processes must first render the animal insensible quickly and painlessly to be considered a humane practice for good animal welfare. Many techniques used to create insensibility or brain death in sturgeon aquaculture do not accomplish this goal due to their size and anatomy. The sturgeon’s cartilaginous skeleton requires greater force to cause brain damage when compared to bony fish. Current techniques for cranial concussion utilize a bat, fish priest or another blunt instrument, or a non-penetrating captive bolt (NPCB). While NPCB provides superior reliability in both placement location and force, there are no studies to date describing the appropriate force required to ensure brain death in sturgeon. This study aims to provide the appropriate NPCB operating pressure (PSI) for juvenile meat fish and adult caviar-ready white sturgeon. Adult caviar-ready, 6-9 yr old female white sturgeon, (mean mass = 27.1kg) and 2-3 yr old juvenile male white sturgeon (mean mass = 8.4kg) were stunned with a Jarvis HPS-1 pneumatic NPCB with a 2.5 cm striking surface. The caviar-ready sturgeon were slaughtered on a commercial farm using 175, 200, and 225 PSI delivered by compressed nitrogen gas monitored by a regulator. Control adult sturgeon experienced multiple strikes between 120 - 145 PSI by a commercial electric air compressor which is the current farm practice. All fish were exsanguinated as a second step termination method. Juvenile sturgeon were stunned at 120, 145, and 170 PSI. Skull and brain were removed by a reciprocating saw, fixed in 37% formaldehyde, and histology of the brain was performed to quantify tissue damage. This study validates the use of NPCB as an effective tool as part of a humane slaughter protocol when operated at specified settings for sturgeon, and improves upon currently available techniques and fish welfare for all cartilaginous fish.