Heart-cutting two-dimensional liquid chromatography methods for quantification of 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole in Class III caramel colours

Heart-cutting two-dimensional liquid chromatography/UV detection methods were developed to determine 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole (noted as THI) in Class III caramel colours. In the first dimension, the caramel sample diluted in water was fractionated either by reversed-phase chromatography on C18 silica packing or by ion-exchange chromatography using strong cationic exchanger (SCX) stationary phase. In each case, THI elution domain was recovered in a loop and transferred to a second column packed with porous graphitic carbon (PGC) for the selective separation of THI. From the accuracy profiles with acceptance limit fixed at 20% and β-expectation tolerance interval fixed at 90%, the two methods, C18–PGC and SCX–PGC, were validated for THI/caramel ratios included between 5–50 and 10–50 ppm, respectively. Finally, the two methods were compared for the analysis of numerous caramel samples.