Chemical restraint of large felids is necessary for the handling, transportation, clinical assessment, and induction of anesthesia. The objective of this study was to evaluate the efficacy and safety of chemical restraint using a combination of tiletamine, zolazepam, detomidine, and atropine with interspecific allometric scaling doses in captive Panthera onca. There were ten males (two melanistic ones) and five females weighing between 35 and 80 kg. The evaluation of the chemical restraint protocol was carried out during routine management of the animals for physical examination, collection of blood and bone marrow samples, dental evaluation, and other procedures. During pre-established intervals, rectal temperature, heart rate, respiratory rate, nociception of thoracic and pelvic limbs, muscle relaxation and loss of the righting reflex were monitored. Loss of the righting reflex occurred at 8 ± 1.89 minutes post-injection (MPI). Respiratory rate returned at 77.6 ± 8.18 MPI; return of consciousness occurred at 112.6 ± 12.28 MPI, and complete recovery at 153 ± 12.47 MPI. There were no statistical differences between heart and respiratory rate means and oxyhemoglobin saturation. For rectal temperature, there was a significant reduction. The protocol analyzed was efficient for the chemical restraint of jaguars, enabling the manipulation, transportation, physical examination, dental evaluation, blood and bone marrow sample collection, and other minor invasive procedures of medium duration in these animals.