The fatty acid pattern of serum phospholipids, cholesteryl esters, triglycerides and free fatty acids was measured before and after a 5-h two-step euglycemic hyperinsulinemic clamp (75 and 1400 microU/ml) in 21 non-insulin-dependent diabetics and 14 age-, weight-, and sex-matched healthy controls. Acute hyperinsulinemia was associated with a statistically significant increase in essential fatty acid and a decrease in non-essential fatty acid contents in triglycerides while the levels of serum triglycerides and free fatty acids dropped in both groups. The fatty acid composition of phospholipids and cholesteryl esters remained unchanged as did the levels of serum phospholipids, total cholesterol and HDL cholesterol.