The aim of this work was to clarify the formation of ethyl carbamate (EC) and its influence factors throughout the production process of Cantonese soy sauce. The results showed that EC was not detected in the koji-making and early moromi fermentation stages, but started to be generated when pH of the moromi decreased to about 4.9—at the same time, the levels of ethanol, urea and citrulline increased significantly. Most EC was formed during raw soy sauce hot extraction (40.6%) and sterilization (42.9%) stages. The EC content exhibited the highest correlation with ethanol throughout the whole production process (R = 0.97). The simulation soy sauce produced in laboratory led the same conclusion—moreover, the contents of EC, ethanol and citrulline were higher in soy sauce fermented at 30 °C than in soy sauce fermented at 15 °C. Extraction of raw soy sauce by squeezing contributed little to EC formation. Further research showed that citrulline and ethanol led to significant increases in EC levels in raw soy sauce upon heating. These results indicate that ethanol and citrulline are two critical precursors of EC and that EC is mainly formed during the heat treatment stage of soy sauce.

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