The gross behavior of excitatory amino acids in patients with traumatic brain injury (TBI), including uptake, transport, metabolism, and clearance, was investigated by analysis of the levels of 41 amino acids in the blood of the jugular vein (JV), which is the primary venous drainage conduit of the brain, and a peripheral artery. Blood samples from the JV and a peripheral artery of eight patients with TBI were collected at 6 h, 6 to 24 h, and over 24 h after TBI, and analyzed using high performance liquid chromatography. Blood samples from 101 normal subjects were also measured. The levels of glutamate (Glu), gamma-aminobutyric acid (GABA), aspartate, glutamine, and cystine deviated from the normal range, and were considered pathological. The level of Glu in the JV was significantly lower than that in the artery (p < 0.05), and the level of GABA in the JV was significantly higher than that in the artery (p < 0.01), but the other three amino acids showed no significant differences. Significantly chronological changes in the difference between the blood levels in the JV and artery were observed for Glu. Measurement of the Glu level in the JV and artery may indicate gross metabolic change in the brain following TBI.