A method for the assay of free sulfate and total sulfoesters in human breast milk by ion chromatography is described. After incubation in 1 M hydrochloric acid at 95 degrees C for 90 min, hydrolytic cleavage of sulfoester standards was essentially complete. The increase in free sulfate after hydrolysis was used as a measure of total acid-labile sulfoesters. We found that this fraction [222 +/- 16 mumol/l (mean +/- standard error), n = 29] comprised 87% of the total sulfate in mature milk. Free sulfate (35 +/- 3 mumol/l) therefore makes only a small contribution to the total sulfate pool available to human infants.