Abstract <p>High-performance liquid chromatography-tandem triple quadrupole mass spectrometry was validated to analyze acrylamide in coffee samples. Under optimal conditions, the quantitative range for acrylamide was 50–1000 µg/L, and the method detection limit was 30 µg/L. The recovery ranged from 85.2 to 97.3%, with high repeatability and reproducibility. The measurement uncertainty of the method was calculated using a top-down approach and determined to be 12.3%. The validated method was then applied to analyze 68 samples of roasted coffee beans collected from markets in Ho Chi Minh City. The results showed that 51.5% of the samples contained acrylamide at concentrations ranging from 51 to 560 µg/kg, and 10.3% had acrylamide contamination exceeding the maximum acceptance level. The results also demonstrated that the acrylamide contamination level in medium-roast coffee was much higher than that in light-roast and dark-roast coffee. The observation indicates that the level of acrylamide contamination of roasted coffee sold in Ho Chi Minh City markets is an excellent concern for food safety and quality management.</p>