A thin dialysis tube was implanted stereotaxically under halothane anesthesia in the caudate nucleus of Sprague-Dawley rats and perfused with Ringer solution at a rate of 2 microliters/min. Initially there was a high rate of purine outflow but after 1-2 h of perfusion the rate was essentially constant (anesthetized - adenosine 0.4 +/- 0.04 microM, inosine 0.8 +/- 0.2 microM; non-anesthetized - adenosine 0.33 +/- 0.03 microM, inosine 0.21 +/- 0.07 microM). Hypoxia (9% O2) increased the levels more than 3-fold. The adenosine deaminase inhibitor erythro-2-(2-hydroxy-3-nonyl)adenine (EHNA) increased the adenosine level and decreased the inosine level. In vitro recovery of adenosine was about 30%. Therefore, we conclude that the free exchangable concentration of adenosine in the rat brain is likely to be 102 micro M. This level is high enough to potentially affect central nervous function.