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Background: Repeated exposure to elevated postprandial triglycerides, as seen with typical Western diets, contributes to atherosclerosis and vascular disease. We determined if a single serving of non-fat milk added to a high fat tolerance test could attenuate postprandial hypertriglyceridemia in individuals who consume a high fat diet. Methods: In this placebo-controlled, randomized, crossover experimental study, 30 overweight/obese adults consumed a high-fat tolerance test meal combined with either non-fat milk, carbohydrate control drink, or caloric control drink. Results: Plasma triglycerides increased over time with no significant differences between interventions. Peak plasma triglyceride levels during HFTT were significantly related to dietary fat intake (r = 0.30, p < 0.05). When participants were divided into tertiles based on habitual dietary fat intake, the higher fat diet group exhibited reduced triglyceride net integrated area under the curve when supplemented with non-fat milk. No significant differences in hemodynamic measures (brachial flow-mediated dilation and femoral vascular conductance) were observed between the milk and caloric control trials for either the low fat or high fat diet groups. Conclusions: A single serving of non-fat milk may attenuate acute hypertriglyceridemia in individuals who chronically consume a high fat diet, offering a simple and easily implemented option for managing elevations in postprandial triglycerides. Keywords: Milk, Dairy products, Postprandial lipemia, Triglycerides, Hemodynamics, Flow-mediated dilation