Evidence for diet quality representing a modifiable risk factor for age-related cognitive decline and mood disturbances has typically come from retrospective, cross-sectional analyses. Here a diet screening tool (DST) was used to categorize healthy middle-aged volunteers (n = 141, 40–65 years) into “optimal” or “sub-optimal” diet groups to investigate cross-sectional associations between diet quality, cognitive function, and mood. The DST distinguished levels of nutrient intake as assessed by Automated Self-Administered 24-h dietary recall and nutrient status, as assessed by blood biomarker measures. Compared with the “sub-optimal” group, the “optimal” diet group showed significantly higher intake of vitamin E (p = 0.007), magnesium (p = 0.001), zinc (p = 0.043) and fiber (p = 0.015), higher circulating levels of vitamin B6 (p = 0.030) and red blood cell folate (p = 0.026) and lower saturated fatty acids (p = 0.012). Regarding psychological outcomes, the “optimal” diet group had significantly better Stroop processing than those with a “sub-optimal” diet (p = 0.013). Regression analysis revealed that higher DST scores were associated with fewer mood disturbances (p = 0.002) and lower perceived stress (p = 0.031), although these differences were not significant when comparing “optimal” versus “sub-optimal” as discrete groups. This study demonstrates the potential of a 20-item diet screen to identify both nutritional and psychological status in an Australian setting.