In this study, a novel method using gas chromatography–mass spectrometry coupled with ethanol and K2HPO4 aqueous two-phase system (ATPS) was established for the quantitative determination of trace ethyl carbamate (EC) in red wine. The parameters that influence EC extraction in an aqueous two-phase system, including extraction temperature, time, pH, and ethanol concentration, were optimized. Method validation results indicated that the regression coefficient of the proposed method was 0.9979 in the linear range of 10 to 100 μg/L, and the limits of detection and quantification were 2.8 and 9.2 μg/L, respectively. Four red wine samples made from different grape varieties were processed by the proposed method for the repeatability verification, and EC concentrations were between 15.8 and 37.3 μg/L, with the relative standard deviations ranging from 3.5 to 6.6%. Results of the precision assay showed the average recovery of EC in red wine at 95.4 to 107.1%, with the relative standard deviations ranging from 1.4 to 6.2%. This method proved to be simple and reliable for quantitative determination of trace EC in red wine and would give guidance for quality monitoring of various red wines in the production process.
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