Thirty-nine reptiles representing 11 squamate species were used to examine the effects of intravenous alfaxalone. All animals examined were captive-born, healthy, and had no history of chronic illness. Alfaxalone at a dose of 9 mg/kg was injected into the ventral coccygeal vein of each animal, and heart rate, respiratory rate, cloacal temperature, time from loss to regaining righting reflex, spontaneous movement, and response to noxious stimuli were recorded. Intravenous alfaxalone at a dose of 9 mg/kg was an effective sedative in most species examined with no obvious complications observed. Loss of righting reflex was not achieved in four (57.1%) blotched bluetongue lizards (Tiliqua nigrolutea), two (33.3%) coastal carpet pythons (Morelia spilota mcdowelli), and one (50%) red-bellied black snake (Pseudechis porphyriacus). Further research is required for alfaxalone use in these species.