The content of luteolin in the different parts of Lithospermum erythrorhizon was determined by high performance liquid chromatography (HPLC). Luteolin was quantified by a reverse-phase column with gradient solvent program (water : ACN = 90 : 10 to 77 : 23 for 60 min). UV detection was conducted at 330 nm. The content of luteolin was measured in the seeds (0.28 mg/g), stems (0.30 mg/g), and roots (0.16 mg/g) of L. erythrorhizon. The presence of luteolin in Lithospermum species is important in agricultural crop production for increasing the amounts of clinically available medicine and health supplements.