We have inhibited the in vitro generation of non-esterified fatty acids in plasma samples taken during haemodialysis by cooling the samples rapidly, centrifuging and freezing within 5 min of sampling and completing the assay on the same day. The relationship of non-esterified fatty acid concentrations during haemodialysis to the acetate concentration of the dialysate and the profile of circulating lipase activities was studied. With this sample-handling protocol we found lower non-esterified fatty acid concentrations than reported in many other studies and the results suggest that, under normal conditions of haemodialysis, the circulating non-esterified fatty acid concentrations and the associated risk of unwanted side-effects may have been previously over-estimated due to artefactual in vitro sample lipolysis. We also showed that, at the end of dialysis, when circulating lipase activities were least, the non-esterified fatty acid concentrations were lower (P < 0.05) with an acetate dialysate containing 37 mmol/l acetate than with a bicarbonate dialysate containing 3 mmol/l acetate.