There are many studies on vitamin B6 (pyrodoxine) or tryptophan (Trp) as a way to increase mood but there are little to no studies with these two nutrients supplemented together. Trp is the precursor to serotonin that requires the cofactor pyridoxal phosphate (PLP). Serotonin plays a role in mood, sleep, appetite, and other wellbeing aspects and it is believed that low levels of serotonin is associated with the risk of developing depression, anxiety, and bipolar disorders. The amount of free Trp that can pass the blood-brain barrier (BBB) is influenced by factors such as cortisol, insulin, and competition from branch chain amino acids (BCAA). College students who exercise on a regular basis and participate in club sports may experience higher cortisol levels from stress of college and higher physical activity. Cortisol decreases the Trp levels in the plasma while BCAAs compete with Trp to pass through the BBB. Insulin promotes the passage of free Trp through the BBB. In the present study, 28 healthy active college students (21.0 ± 2.1 years, 24.5± 3.1 kg/m2) were divided into three groups: vitamin B6 (n=11), Trp (n=10), or both (n=10) (2 did not complete study). Blood serum pyridoxine levels and mood states were measured at baseline and at 4 weeks with Profile of Mood States (POMS), Depression Anxiety Stress Survey (DASS), Life Orientation Test-Revised (LOT-R), and Epworth sleep scale. In the combined sample, the total POMS score improved during the study (p=0.039) and the total DASS score tended to improve during the study (p=0.068). Thus, mean depression scores for all participants decreased during the 4-week supplementation study. However, there were no time x treatment effects noted at study completion. At baseline 18% of the participants were marginally deficient in vitamin B6 (serum pyridoxine <30nmol/L), and their total POMS score was raised 78% in comparison to participants with adequate vitamin B6 status (p=0.08). DASS scores were raised 48% in vitamin B6 deficient participants versus those with adequate vitamin B6 status (p=0.243). There were no significant changes (time or time x treatment) during the course of the study for the LOT-R or sleep scores. In summary, vitamin B6 deficiency in college student athletes was remarkably high (18%) compared to the national average reported by the CDC in 2012 (10.5%), and participants with vitamin B6 deficiency displayed heightened unfavorable mood states. Moreover, supplementation with vitamin B6, tryptophan, or vitamin B6 and tryptophan improved mood state in college student athletes, but there were no differences between treatments.