In greenhouse experiments, the contents of nitrates in headed lettuce harvested in spring (in two terms with a 5-day delay) and in autumn at two day-times (in the morning and in the afternoon) were compared. Effects of nitrogen level in the substrate and nitrogen form, as well as potassium fertilizer type and its level in the substrate were identified. Content of nitrates in fresh matter of plants was determined by colorimetric method using direct cadmium reduction and Griess reagent. Smaller amount of nitrates in lettuce leaves was obtained using reduced nitrate forms and potassium chloride fertilizers. Increase of potassium content in the substrate contributed to an increase of nitrate content in plants. Significant decrease of nitrogen content, particularly in spring, was found when lettuce was harvested in the afternoon (at 3.00 pm) as compared with morning harvest (at 7.00 am). In the phase of consumption maturity, the delay of harvest by 5 days in spring significantly decreased (on average by 56%) the nitrate content in lettuce.