Lysophospholipids (LPL), a new feed additive, were supplemented to a pelleted total mixed ration (TMR) of dairy cows to examine its effects on feed intake, production, and composition of milk and plasma biochemical parameters. Two dietary treatments included diets supplemented without (control diet; CON) or with LPL at a dose of 0.5 g/kg of pelleted TMR. Twelve multiparous, mid-lactation, Holstein cows (Bodyweight 730 ± 9.3 kg; 100 ± 6.0 days in milk) were randomly assigned to one of the two dietary treatments with a 42-day measurement period after a 14-day adaptation period. Feed and water were provided ad libitum. Feed intake and milk yields were recorded daily, blood samples were collected fortnightly, and milk samples weekly. The results showed that the supplementation of LPL did not change feed dry matter intake, milk yields, and milk composition. However, it increased total protein and globulin and the activity of alkaline phosphatase and decreased total cholesterol in plasma. This study suggests that LPL may have beneficent effects in animal health but might be not a feasible feed additive to increase production for dairy cows fed a pelleted TMR.