Vitamin C or L-ascorbate is an effective endogenous reducing agent influencing the functions that are involved with the redox mechanism. Detection of oxygen tension changes by the carotid body chemoreceptors is one such function. In this study we investigated the hypothesis that the level of ascorbate, if present, could change in the carotid body as a result of ascorbate's interaction in the chemosensing process, which would be reflected in the production of the ascorbyl radical. We addressed this issue by examining changes of the ascorbyl radical, using the in vitro electron spin resonance spectroscopy (ESR), in the carotid bodies dissected from pentobarbitone anesthetized cats exposed in vivo to three eucapnic, breathing gas mixtures of different O2 concentration: normoxic, 21% O2; hypoxic, 7% O2; and hypoxic-reoxygenated, 7% O2 followed by 100% O2. Each group consisted of five cats, yielding five pairs of carotid bodies for the ESR signal recording. We found that the intensity of ESR signals, measured as peak-to-peak amplitude, was diminished by 39 and 43% in the hypoxic and hypoxic-reoxygenated groups, respectively, compared with that in normoxia. The study shows that ascorbate was present in the normoxic carotid body and was depleted by hypoxia. We conclude that ascorbate is part of free radical mechanisms operative in the carotid body in hypoxia.