Cortisol awakening response (CAR) refers to the dynamic change of cortisol concentration within 1 h after awakening. Trait anxiety is a general risk marker of anxiety disorders. The present study aimed to investigate the effect of trait anxiety on CAR using R30 (change in cortisol level at 30 min after awakening) and AUCi (the area under the curve with respect to the increase) as indicators. 133 college students were divided into high trait anxiety (HTA) and low trait anxiety (LTA) group according to the median score of the trait version of State-Trait Anxiety Inventory. Saliva samples were collected immediately upon awakening, 30 min, 45 and 60 min on two consecutive mornings. The results showed that, (1) decreased CAR was found for the male than female participants. (2) Compared to the LTA group, the HTA group showed decreased R30 and AUCi. (3) Both R30 and AUCi were negatively correlated with trait anxiety scores. These results demonstrated that trait anxiety might weaken the CAR, and both R30 and AUCi can be used as CAR indicators in detecting trait anxiety. Future research on CAR should consider the moderating effect of trait anxiety.