The in vivo formation of dimethyltryptamine was studied in rabbits, rats and monkeys. When C14-labelled N-methyltryptamine was administered by intravenous injection to rabbits, C14-dimethyltryptamine was found in lung, the principle site of the methyltransferase that biosynthesizes this psychotogen. Unequivocal evidence for C14-dimethyltryptamine formation in rat tissues was not obtained. When rabbits were given non-radioactive N-methyltryptamine intravenously, dimethyltryptamine appeared in carotid arterail blood, peaking within the first minute after injection of the precursor. USing this assay procedure we could not demonstrate dimethyltryptamine synthesis in the rhesus monkey.