Purpose: We present a case report about an acute intoxication episode after an oral administration of Ayahuasca and dermal exposure of Kambo for treatment of depression. The clinical features observed were hallucination, agitation, tremors of extremities, oral paresthesia, skin lesions and seizures. Diazepam was administered by the emergency service and was effective in controlling hallucination, but failed to control agitation and seizures. Methods: Patient biological fluids (urine and serum) and the samples of Ayahuasca and Kambo were submitted to toxicological analysis using liquid-liquid extraction followed by liquid chromatography–tandem mass spectrometry and high-resolution electrospray ionization-mass spectrometry. Results: The main active compounds present in Ayahuasca, N,N-dimethyltryptamine, harmine, harmaline and tetrahydroharmine were found and quantified in the different samples, confirming the use by the patient. In Kambo secretion used in the ritual, we were able to find sixteen potently active peptides: adenoregulin, bombesin, bombesinnona peptide, bradykinin -phe(8)-psi-CH2NH-arg(9)-, caerulein, deltorphin, neurokinin B, phyllomedusin, phyllocaerulein, phyllokinin, phyllolitorin, preprotachykinin B (50–79), ranatachykinin A, sauvagine, T-kinin and urechistachykinin II. Conclusions: The patient was discharged the day after exposure without any sequel. Clinical and toxicological analysis indicated that the symptoms presented by the patient occurred due to a joint action produced by the substances identified in both materials. To the best of our knowledge, this is the first case involving probable intoxication by simultaneous administration of Ayahuasca and Kambo.