While the formalin test is a widely used behavioral model of tonic chemogenic pain, little is known about the responses of primary afferent nociceptors to formalin. Formalin (2.5%, 50 μl) was injected either directly in or adjacent to the mechanical receptive fields of single C-fibers isolated from the saphenous nerve of pentobarbital-anesthetized rats. The average formalin-evoked response in C-fibers (n = 29) over time was biphasic. This biphasic time course of the C-fiber response to formalin is similar to that of the behavioral response in the awake animal and is compatible with the hypothesis that increased C-fiber activity contributes to the behavioral response in phase 2, as well as in phase 1 of the formalin test.