Determination of 4-Methylimidazole and 2-Acetyl-4(5)- tetrahydroxybutylimidazole in Caramel Color and Processed Foods by LC-MS/MS
In this study, the quick HPLC-MS/MS method for the simultaneous separation of 2-acetyl-4(5)-tetrahydrox-ybuthylimidazole (THI) and 4-(5)-methylimidazole (4-MI) in alkaline medium was used for caramel color and processed foods in Korea. After a simple sample pretreatment, 51 4-MI-labeled samples were positive for 4-MI and 2 also contained THI. The concentration of 4-MI was 260.5 ~ 24,499.3 μg/kg in caramel color, less than LOD ~ 1,712.5 μg/kg in sauce, 1,242.3, 5,972.2 μg/kg in balsamic vinegar, 2,118.3 ~ 5,802.4 μg/kg in complex seasoning, 82.7 ~ 5,110.6 μg/kg in curry, and 29.9 ~ 464.4 μg/kg in soft drinks. The recovery rate of 4-MI was 97.1 ~ 111.0% in sauce and 81.9 ~ 110.0% in powder and that of THI was 83.6 ~ 106.4% in sauce and 61.2 ~ 99.4% in powder. Our results concluded a safe amount of 4-MI and THI compared to the limit of Korea additive code but the processed foods do not have a limit of caramel color and 4-MI in Korea. Therefore, research and monitoring of 4-MI and THI is needed for processed foods in Korea.
Determination of 4(5)-methylimidazole in soy sauce and other foods by LC-MS/MS after solid-phase extraction.
A method for the determination of 4(5)-methylimidazole (4MeI) in naturally brewed soy sauce was developed for the first time using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). SPE on silica-based reversed-phase cartridges with heptafluorobutyric acid as an ion-pairing reagent was used for the efficient cleanup of 4MeI. A multimode ODS column was employed for the chromatographic separation. To subtract the matrix effect during LC-MS/MS analysis, a standard addition method was used. The levels of 4MeI found in naturally brewed soy sauce were extremely low (ranging from <0.002 to 0.023 μg/g), whereas those in soy sauces containing caramel color were generally high (ranging from 0.43 to 4.8 μg/g). The method proved to be useful for the analysis of 4MeI in other foods such as caramel colors, drinks, and Worcestershire sauce.
Etiology of ammoniated hay toxicosis.
Some animals consuming hay treated with anhydrous ammonia have developed neurological signs including hyperexcitability, circling and convulsions. A series of experiments was conducted to identify tentatively the toxin and determine its mode of action. Three out of four sheep fed ammoniated orchardgrass hay (approximately 4% ammonia on a dry basis) developed convulsions. Two of the three sheep died within 18 h of the onset of signs. The concentrations of blood lactate and pyruvate were elevated in the symptomatic sheep (P less than .05). A proposed toxin, 4-methyl imidazole, did not induce the syndrome when 750 mg/d (approximately 10 times the dietary amount) were administered orally. Four out of five calves that received milk from cows fed ammoniated oat hay (approximately 5% ammonia on dry basis) displayed hyperexcitability and circling. Concentrations of blood lactate and pyruvate were also elevated in the calves. The crude alkaloid fraction of the toxic milk produced neurological signs similar to those of the calves when injected into mice. A fluorescent compound was found in the alkaloid fraction of toxic milk and ammoniated hay, but not in control milk or untreated hay. The fluorescent compound was quite labile; hence, characterization has been unsuccessful thus far.