This study investigated salivary cholesterol of children from 6 to 16 years of age in response to dietary intervention. One thousand sixty-two infants started in the prospective, randomized project. At 3 years of age, every fifth child was invited into the study (n=178). Of these, 148 enrolled, and 86 completed the oral sub-study at 16 years of age. The intervention aimed at restricting the child's saturated fat and cholesterol intake. Control children received no special recommendations. Every third year, paraffin-stimulated saliva samples (10.0 mL) were collected for cholesterol assays. Nutrient intakes and serum total cholesterol concentrations were regularly followed up by means of 4-day food records and blood samples. Intake of saturated fatty acids (SAFA) was lower in the intervention than in the control group (p<0.001). Salivary cholesterol concentration increased from 1.9 (±1.1) µmol/L at 6 years of age to 16.0 (±9.0) µmol/L at 16 years of age. The increase was smaller in the intervention than in the control group (p<0.001). The ratios of salivary to serum cholesterol concentrations tended to be higher in boys than in girls (p=0.07). Thus, dietary intervention was reflected in children's salivary cholesterol values more sensitively than in serum cholesterol values. (clinicaltrials.gov NCT00223600).