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The influence of microemulsion electrokinetic chromatography (MEEKC) operating conditions, such as the type of water-immiscible alcohol, aqueous phosphate buffer concentration, pH, as well as the addition of methanol and 2-propanol, on acrylamide migration has been studied. These parameters have been optimized taking into account the presence of matrix signals, in order to avoid the interference of these peaks in acrylamide determination. The best separations were achieved using a microemulsion consisting of 0.8% m/v n-amyl alcohol, 3.3% m/v sodium dodecyl sulfate (SDS), 6.6% m/v 1-butanol, and 89.3% m/v 40 mM phosphate buffer at pH 6.5 working at 15 kV in uncoated silica capillaries. Linear calibration curves over the range studied (1.25-125 microg x mL(-1)), the detection limit (0.70 microg x mL(-1)), and both run-to-run (up to 3.4% for concentration and 1.6% for time values) and day-to-day precision (lower than 11.6% for concentration) have been established. Finally, the applicability of the MEEKC method developed has been demonstrated by analyzing levels of acrylamide present in samples of home-made French fries.