Detailed information on the anaesthetic and cardiorespiratory effects of drug combinations used for the chemical immobilization of Kinkajous (Potos flavus) is scarce. This study assessed the effects of ketamine (2.5mg/kg), xylazine (1mg/kg) and midazolam (0.5mg/kg) combination in P. flavus. Five clinically healthy adult Kinkajous of both sexes were included. Heart rate, respiratory rate, oxygen saturation, blood pressure and body temperature were recorded at five-minute intervals for 25 minutes. Then, animals received 0.125mg/kg of yohimbine by intramuscular injection. Anaesthetic depth was assessed based on stimulus response and muscle tone. Induction, immobilization, and recovery periods were recorded and qualitatively assessed based on the absence of adverse effects. The durations of the induction, immobilization, and recovery periods were 9.42±1.73, 33.33±2.16, and 31.37±5.82 minutes. All periods showed good quality and adequate anaesthetic depth was achieved. Mean heart and respiratory rates were 99±20 beats/minute and 44±9 breaths/minute. Both parameters decreased over the duration of the anaesthesia but they did not reach levels suggesting either bradycardia or bradypnea. Mean body temperature was 37.1±1.5 °C and it also showed a decreasing trend over the duration of the anaesthesia. Mean oxygen saturation was 92±6% and it showed a mildly increasing trend over the duration of the anesthesia. Mean blood pressure was 129±23 mmHg and mild to moderate hypertension was observed. No mortality occurred and no adverse effects were observed in any of the individuals during the three months following immobilization. The assessed anaesthetic combination effectively immobilized the P. flavus individuals, provided good quality and acceptable duration of both induction and recovery periods. It should, however, not be used in Kinkajous with either known hypertension record or pre-existing target organ disease (e.g., renal failure, retinopathy).