Simultaneous quantitation of 2-acetyl-4-tetrahydroxybutylimidazole, 2- and 4-methylimidazoles, and 5-hydroxymethylfurfural in beverages by ultrahigh-performance liquid chromatography-tandem mass spectrometry.
An ultrahigh-performance liquid chromatography (UHPLC) tandem mass spectrometric (MS/MS) method was developed for the simultaneous quantification of 2-acetyl-4-tetrahydroxybutylimidazole (THI), 2- and 4-methylimidazoles (2-MI and 4-MI), and 5-hydroxymethylfurfural (HMF) in beverage samples. A C30 reversed-phase column was used in this method, providing sufficient retention and total resolution for all targeted analytes, with an MS/MS instrument operated in selected reaction monitoring (SRM) mode for sensitive and selective detection using isotope-labeled 4-methyl-d(3)-imidazole (4-MI-d(3)) as the internal standard (IS). This method demonstrates lower limit of quantification (LLOQ) at 1 ng/mL and coefficient of determination (r(2)) >0.999 for each analyte with a calibration range established from 1 to 500 ng/mL. This method also demonstrates excellent quantification accuracy (84.6-105% at 5 ng/mL, n = 7), precision (RSD < 7% at 5 ng/mL, n = 7), and recovery (88.8-99.5% at 10, 100, and 200 ng/mL, n = 3). Seventeen carbonated beverage samples were tested (n  = 2) in this study including 13 dark-colored beverage samples with different flavors and varieties and 4 light-colored beverage samples. Three target analytes were quantified in these samples with concentrations in the range from 284 to 644 ng/mL for 4-MI and from 706 to 4940 ng/mL for HMF. THI was detected in only one sample at 6.35 ng/mL.
Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, California 94086, United States. leo.wang@thermofisher.com
Jinyuan
J
Wang
William C
WC
Schnute
D002253:Carbonated Beverages / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005505:Food Coloring Agents / Q000032:analysis; D005506:Food Contamination / Q000032:analysis*; D005662:Furaldehyde / Q000031:analogs & derivatives* / Q000032:analysis; D007093:Imidazoles / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D005505:Food Coloring Agents; D007093:Imidazoles; C008046:5-hydroxymethylfurfural; C049929:2-acetyl-4(5)-tetrahydroxybutylimidazole; D005662:Furaldehyde; C023439:4-methylimidazole; C032655:2-methylimidazole