Analysis of 4(5)-methylimidazole in soy sauce by a quick, easy, cheap, effective, rugged, and safe approach and liquid chromatography-mass spectrometry

Abstract

4(5)-Methylimidazole (4(5)-MI) is a potential carcinogen with low molecule weight, highly polarity, and weak basicity. The traditional way to extract and clean-up 4(5)-MI in soy sauce using solid phase extraction is tedious and time consuming. Here we proposed a method for the determination of 4(5)-MI in soy sauce by combining a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction with liquid chromatography-mass spectrometry analysis. The impacts of solution pH, water addition, and cleanup procedure on 4(5)-MI extraction efficiency were studied. An optimized sample preparation approach involved a single step liquid-liquid extraction between acetonitrile and soy sauce under alkaline conditions, followed by primary and secondary amine clean-up. The analytical method was validated with soy sauce at three spiking levels (10, 50, 500 ng/g). The method recovery (96.2–107%) and intra-day/inter-day precision (4.1–8.4%/6.9–11.7%) were satisfactory. The method quantification limit was 10 ng/g. The developed method was successfully applied for the determination of 4(5)-MI in fourteen commercial soy sauces from local markets. The results obtained in this work suggests that the method is suitable for the analysis of 4(5)-MI at low concentrations in high-salting and protein-containing soy sauce matrix.