We demonstrate the existence of two types of paramagnetic centers in a pure graphene oxide. Saturation features of the electron paramagnetic resonance (EPR) spectrum in the temperature range of 4.2–300 K reveal that one of these centers has the spin–lattice relaxation time longer than 1.6 μs at room temperature. The spectrum of these centers consists of a central line and two satellite lines. The satellite lines result from the forbidden transitions in the hyperfine structure induced by protons in the vicinity of the slowly relaxing centers. The observability of the satellites can be used as a characteristic signature of graphene oxide purity. Since the number of slowly relaxing centers is reduced during chemical reduction of graphene oxide, this type of centers can be attributed to the isolated unfunctionalized carbons in the highly functionalized regions of GO. The second type of paramagnetic centers has shorter relaxation times and dominates in the reduced graphene oxide.