Hydroxyethyl starch (HES) found in rat serum and spleen after single and daily administrations of 0.9 g/kg for 1 week was characterized by gas-liquid chromatography. There was very little difference in the degree of substitution (D.S.) and molar substitution (M.S.) of HES in serum samples obtained at 1 hour and 57 days after multiple doses and of HES in spleen samples obtained at 1 hour and 168 days after a single dose of HES. The small increase in D.S. and M.S. was due to a decrease in the glucose content and not due to a change in the ratio of mono- to poly-substituted glucoses.