4(5)-methylimidazole (4(5)-MI) is carcinogenic, nitrogen-containing compound, mainly found during the manufacturing of caramel coloring. Hence, presence of 4(5)-MI is well-known in any food products with addition of caramel coloring for desirable sensory characteristics, such as cookies. Limited work has been conducted to develop suitable analytical method and to investigate effect of caramel coloring in cookies. This study aimed at developing the analytical method for quantification of 4(5)-MI and confirming influence of caramel coloring on level of 4(5)-Mi in cookies. Gas chromatography-mass spectrometry (GC-MS) was utilized for qualification and quantification of 4(5)-MI. Sample preparation procedure specialized in bakery products was fully developed in this study. The concentration of 4(5)-MI in 15 commercial cookies and biscuits ranged from 71.5 to 1254.8 ng/g. Correlation equation (y = 706.42x + 21.792) was obtained to estimate effect of caramel colorant on level of 4(5)-MI in cookies. Further, analytical method developed and results of correlation equation can be utilized in future studies on reduction of 4(5)-MI in many food.

4(5)-methylimidazole (4(5)-MI) is carcinogenic, nitrogen-containing compound, mainly found during the manufacturing of caramel coloring. Hence, presence of 4(5)-MI is well-known in any food products with addition of caramel coloring for desirable sensory characteristics, such as cookies. Limited work has been conducted to develop suitable analytical method and to investigate effect of caramel coloring in cookies. This study aimed at developing the analytical method for quantification of 4(5)-MI and confirming influence of caramel coloring on level of 4(5)-Mi in cookies. Gas chromatography-mass spectrometry (GC-MS) was utilized for qualification and quantification of 4(5)-MI. Sample preparation procedure specialized in bakery products was fully developed in this study. The concentration of 4(5)-MI in 15 commercial cookies and biscuits ranged from 71.5 to 1254.8 ng/g. Correlation equation (y = 706.42x + 21.792) was obtained to estimate effect of caramel colorant on level of 4(5)-MI in cookies. Further, analytical method developed and results of correlation equation can be utilized in future studies on reduction of 4(5)-MI in many food.