We demonstrate the presence of reticuline, an isoquinoline alkaloid that was purified and identified in the rat brain. This was achieved by high-performance liquid chromatography coupled with electrochemical detection. This material was finally identified by nano-electrospray ionization quadrupole time-of-flight tandem mass spectrometry. The expression of this tetrahydroisoquinoline alkaloid in rat brain is at 12.7+/-5.4 ng/g wet tissue. Furthermore, rat chow, rat small and large intestine and bacteria cultured from these tissues did not contain either morphine or reticuline, eliminating the possibility of contamination or an exogenous source of these compounds. This finding adds information which suggests that morphine biosynthesis may occur in rat neural tissues, and that its biosynthesis pathway may be similar to that reported in the poppy plant.