Interstitial glycerol may be a useful marker for posttraumatic and postischemic membrane phospholipid (PL) breakdown. Degradation of membrane PLs is thought to be triggered by both calcium and reactive oxygen species (ROS)-mediated mechanisms and to be associated with disturbed energy metabolism. In this study, we investigated the temporal changes of interstitial glycerol, lactate, and glucose after traumatic brain injury in the rat and the effect of pretreatment with the free radical spin trap alpha-phenyl-N-tert-butyl nitrone (PBN; 30 mg/kg i.v.). Microdialysate was sampled continuously in 10-min fractions from 1 h before, until 2 h after a cortical contusion injury produced by the weight-drop technique. The maximal concentration of interstitial glycerol (a ninefold increase) was seen 10-30 min after trauma and subsided during the following 2 h, but remained above base line as compared to sham operated animals. Concomitantly, there was an increase in interstitial lactate (fivefold) and a fall in interstitial glucose, indicating a posttraumatic energy perturbation. PBN treatment significantly attenuated the interstitial accumulation of glycerol and lactate. The results support the concept that ROS are involved in posttraumatic membrane PL breakdown and that PBN improves mitochondrial function after CNS injury. Monitoring of interstitial glycerol with microdialysis may be a valuable tool for studies on membrane PL degradation and the efficacy of neuroprotective drugs in acute CNS injury.