Caramel colour and process by-products in foods and beverages: Part I - Development of a UPLC-MS/MS isotope dilution method for determination of 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole (THI), 4-methylimidazole (4-MEI) and 2-methylimidazol (2-MEI)

Food Chem. 2018 Jul 30;255:348-356. doi: 10.1016/j.foodchem.2018.02.039. Epub 2018 Feb 9.


Caramel colours are used by the food industry in a wide range of foods and beverages. During their manufacturing, low molecular weight compounds such as 4-methylimidazole (4-MEI), the structural isomer of 4-MEI, 2-methylimidazole (2-MEI) and 2-acetyl-4-tetrahydroxy-butylimidazole (THI) are generated. The presence of these inevitable by-products of caramel manufacturing can be hazardous to human health. This publication describes an isotope dilution Ultra-High-performance Liquid Chromatography tandem mass spectrometric method (UHPLC-MS/MS) that was developed and validated for the simultaneous quantification of these impurities in both beverages/liquids and foods. A limit of quantification of 5 μg/kg was obtained for 4-MEI and THI. The expanded measurement uncertainty (U; k = 2) for these compounds was below 51% in beverages/liquids and below 56% in foods. As higher measurement uncertainties were obtained for 2-MEI, the developed analytical procedure can only be used in a semi-quantitative way for this compound.

Keywords: 2-Methylimidazole (Pubchem CID:12749); 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole; 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole (Pubchem CID:108037); 2-methylimidazole; 4-Methylimidazole (Pubchem CID:13195); 4-methylimidazole; Belgium; Beverages; Caramel colours; Food; Isotope dilution; Liquid chromatography; Mass spectrometry; Quantification.

MeSH terms

  • Beverages / analysis*
  • Carbohydrates / analysis*
  • Chromatography, Liquid
  • Food Analysis / methods*
  • Food*
  • Imidazoles / analysis*
  • Isotopes / analysis
  • Tandem Mass Spectrometry


  • Carbohydrates
  • Imidazoles
  • Isotopes
  • caramel coloring