Abstract Objectives Previous research indicates that consumption of strawberries may provide benefits for reduction of atherogenic lipoproteins but has not identified an optimal dose. Our objective was to evaluate effects of 2 doses of strawberry powder, approximately equivalent to 1 and 3 servings of strawberries per day, on serum lipoprotein concentrations. Methods Middle-aged adults (n = 40, age 49 ± 1 year) with elevated LDL-C (140 ± 4 mg/dL) and elevated BMI (29.4 ± 0.4 kg/m2) consumed 0 g/d (control), 13 g/d (low-dose), and 40 g/d (high-dose) of freeze-dried strawberry powder in a randomized crossover design (4-week supplementation periods separated by a 2 week compliance break). Fasting blood samples were obtained on two separate days (and averaged) at study-entry baseline and following each supplementation period. Results There were significant main effects of treatment (P ≤ 0.05) for calculated LDL-C, nonHDL-C, and total cholesterol (TC). In post hoc tests, the low-dose resulted in 5% lower LDL-C vs. the high-dose (P = 0.01), 4% lower nonHDL-C vs. control (P = 0.04), and 3% lower TC for the low-dose vs. control and high-dose (P ≤ 0.04). Compared to baseline, low-dose strawberry supplementation also significantly reduced direct LDL-C (−6.8 mg/dL, P = 0.02), but there was not a main effect of treatment. Conclusions Our results suggest that low-dose supplementation with freeze dried strawberry powder, roughly equivalent to one serving of strawberries per day, was superior to high-dose supplementation for improving atherogenic lipoproteins in overweight adults. Funding Sources California Strawberry Commission.