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A liquid chromatography tandem mass spectrometry method (LC-MS/MS) preceded by a rapid and simple QuEChERS-based sample preparation was developed and validated for the determination of acrylamide in non-centrifugal sugar (NCS), a solid product obtained from sugarcane juice without centrifugation, up to a concentration greater than 90 ºBrix. Adequate linearity in calibration curves, analytical selectivity and sensitivity were achieved. A limit of detection (LOD) and a limit of quantitation (LOQ) of 10 and 20 µg kg−1, respectively, were obtained. Recoveries (84.3–107.5%) and coefficients of variation (CV<15%) for repeatability and reproducibility conditions were appropriate in intraday and interday laboratory conditions. The concentration of acrylamide was determined in 76 commercial samples of NCS from different Latin American countries and ranged from <20 to 1414 µg kg−1. The highest mean acrylamide amounts were found in samples from Peru, Ecuador and Colombia (636, 446 and 401 µg kg−1, respectively). Considering the form of the product, the highest average levels were found in granulated NCS (517 µg kg−1) when compared to block (294 µg kg−1). Within the Colombian samples, organic NCS, made with natural flocculant, presented a higher acrylamide mean concentration (721 µg kg−1) than conventional samples (363 µg kg−1). Although the Margin of Exposure (MOE) values for neurotoxicity suggested that the estimated intakes in Brazil and Colombia are not a concern, MOEs obtained for neoplastic effects raised attention. [ABSTRACT FROM AUTHOR]