Background: Accumulating evidence implicates diet quality in childhood as playing a significant role in adult cardiometabolic health. Polyunsaturated fatty acids (PUFAs) of the n-6 (ω-6) and n-3 (ω-3) series contribute unique protective effects against cardiometabolic disease. As such, the ratio between n-6 and n-3 PUFAs is a dietary metric of interest in the early life span, although an optimum intake ratio has yet to be determined. Objective: This cross-sectional study assesses relations between the ratio of total n-6:n-3 PUFA intake and cardiometabolic risk factors in a racially diverse sample of children (n = 191) from the Admixture Mapping of Ethnic and Racial Insulin Complex Outcomes (AMERICO) study. Methods: Outcome measures included waist circumference, lipid concentrations, fasting glucose, and two 24-h dietary recalls from boys and girls aged 7-12 y who self-reported as European American (n = 81), African American (n = 55), or Hispanic American (n = 55). Linear regression analyses were used to assess associations between predictors of interest and outcomes after adjusting for covariates. Results: PUFA intake reflected in the n-6:n-3 ratio was inversely associated with concentrations of total and LDL cholesterol [β ± SE: -0.359 ± 0.107 (P = 0.001) and -0.189 ± 0.069 (P = 0.007), respectively]. Exploratory analyses showed that the intake of total n-6 PUFAs was not significantly predictive of any cardiometabolic risk factor assessed, whereas total n-3 PUFA intake was positively associated with concentrations of HDL cholesterol (β ± SE: 0.114 ± 0.042; P = 0.007). Conclusions: Results suggest that the effect of n-6 and n-3 PUFA intake reflected in the ratio may be largely driven by n-3 PUFAs in reducing 2 lipid cardiometabolic risk factors among this multiethnic cohort of children. Until an ideal intake ratio is determined, nutritional counseling should focus on meeting recommended levels of both n-3 and n-6 PUFAs in order to establish beneficial childhood dietary patterns that may positively influence adult cardiometabolic health.