It is unknown how consumption of bitter foods and beverages in the maternal diet influences sensory properties of fresh human milk. The aims of this study were (1) to determine the sensory characteristics of fresh human fore and hind milk, (2) to establish relationships between sensory properties and composition of fresh human milk, and (3) to assess the relationship between bitterness of the maternal diet and human milk. Twenty-two lactating mothers generated sensory terms to describe perception of their milk and received training on sensory attribute intensity rating. Mothers kept a 24-h food diary followed by a sensory self-assessment of their fore and hind milks. The odor of human fresh milk was described as neutral, creamy, and sweet, taste as sweet and bitter, and mouthfeel as thin, watery, smooth, and fatty. Sweetness was equivalent to 1.53 g of sucrose/100 mL and was not significantly different between fore and hind milk. Fore milk was significantly less creamy, less fatty, thinner, more watery, and lower in vanilla flavor intensity than hind milk. Carbohydrate content of human milk was positively correlated with sweetness and glutamic acid content with umami. The bitterness of the diet consumed 24 h before lactation was moderately positively correlated with bitterness of fore milk, but not hind milk. We conclude that the consumption of bitter foods may influence the bitterness of human fore milk, which may be another way for breastfed children to learn to accept bitter vegetables and, hence, develop healthier food preferences.