{"id":43564,"date":"2020-10-21T14:30:00","date_gmt":"2020-10-21T14:30:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/reliable-hilic-lcmsms-analysis-of-4-methylimidazole-4-mei-on-raptor-fluorophenyl-columns\/"},"modified":"2025-12-09T20:35:45","modified_gmt":"2025-12-09T20:35:45","slug":"reliable-hilic-lcmsms-analysis-of-4-methylimidazole-4-mei-on-raptor-fluorophenyl-columns","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/ja\/articles-ja\/ffss2524\/reliable-hilic-lcmsms-analysis-of-4-methylimidazole-4-mei-on-raptor-fluorophenyl-columns","title":{"rendered":"Reliable HILIC LC-MS\/MS Analysis of 4-Methylimidazole (4-MEI) on Raptor FluoroPhenyl Columns"},"content":{"rendered":"\n<ul class=\"wp-block-list\">\n<li>Increased retention for 4-MEI compared to a C18.<\/li>\n\n\n\n<li>Flexible Raptor FluoroPhenyl columns can be used in both HILIC and reversed-phase modes.<\/li>\n\n\n\n<li>Exceptional reproducibility assures accurate results analysis after analysis.<\/li>\n<\/ul>\n\n\n\n<p>Caramel colorings are used in the food industry to impart a more desirable color to a wide range of foods and beverages. The many classes of colorings are produced in a variety of different ways, and those classes made using ammonia create 4-methylimidazole (4-MEI) as a byproduct. 4-MEI (CAS# 822-36-6) is classified by the International Agency for Research on Cancer (IARC) in Group 2B as possibly carcinogenic to humans [1]. This designation has generated interest in analytical methods that can be used to determine 4-MEI levels in both the colorings themselves to monitor formulations and also in foods and beverages to assess potential exposure. LC analysis of 4-MEI is a common approach, but as a small, polar compound, 4-MEI is difficult to retain using typical reversed-phase conditions and C18 columns.<\/p>\n\n\n\n<p>As shown below, LC analysis of 4-MEI using HILIC mode and a Raptor FluoroPhenyl column is a much better strategy that yields strong retention and good peak shape. The highly reproducible performance and fast 2-minute analysis time (3.5 minute total cycle time) make this an excellent approach for labs needing high-throughput methods. Raptor FluoroPhenyl columns are recommended for LC analysis of 4-MEI in HILIC mode, but they can also be used in reversed-phase mode for other analyses, giving method developers the flexibility to use the column in both modes and then select the mode that performs best for their specific compounds of interest.<\/p>\n\n\n\n<p>[1] International Agency for Research on Cancer, IARC Monographs on the Evaluation of Carcinogenic Risks to Humans.&nbsp;<a href=\"http:\/\/monographs.iarc.fr\/ENG\/Classification\/latest_classif.php\" target=\"_blank\" rel=\"noreferrer noopener\">http:\/\/monographs.iarc.fr\/ENG\/Classification\/latest_classif.php<\/a><\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><\/div><div class='chromatogram-article-inner-full'><div class=\"chromatogram-article-inner\">\n<div class=\"wp-block-custom-chromatogram-article\"><div class=\"wp-block-custom-chromatogram-article\"><div class=\"chromatogram-image wide-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0559.png\" alt=\"Analysis of 4-Methylimidazole on Raptor FluoroPhenyl by LC-MS\/MS\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_FF0559<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12\">\n<thead><tr><th><\/th><th style=\"text-align: left;width: 75px\">Peaks<\/th><th style=\"text-align: center;width: 75px\">t<sub>R<\/sub> (min)<\/th><th style=\"text-align: center;width: 75px\">Precursor Ion<\/th><th style=\"text-align: center;width: 75px\">Product Ion<\/th><\/tr><\/thead>\n<tbody><tr><td class=\"num\">1.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 4-Methylimidazole (4-MEI)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/822-36-6\/4-Methylimidazole\" target=\"_blank\" rel=\"noopener\">4-Methylimidazole (4-MEI)<\/a><\/td><td class=\"oth\">1.19<\/td><td class=\"oth\">83<\/td><td class=\"oth\">56<\/td><\/tr>\n<\/tbody><\/table><\/div><div class=\"chromatogram-conditions\"><h4>Conditions<\/h4><div class=\"conditions-container container-fluid\"><div class=\"row\"><table class=\"conditions col-lg-6 col-12\"><tr><th class=\"conditions_header\" scope=\"row\">Column<\/th><td>Raptor FluoroPhenyl  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9319A52?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_FF0559\" rel=\"noopener\">cat.# 9319A52<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Dimensions:<\/th><td>50 mm x 2.1 mm ID<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Particle Size:<\/th><td>2.7 \u00b5m<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Temp.:<\/th><td>35 \u00b0C<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Standard\/Sample<\/th><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Diluent:<\/th><td>Acetonitrile<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>100 ng\/mL<\/td><\/tr><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Inj. Vol.:<\/th><td>5 \u00b5L <\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Mobile Phase<\/th><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">A:<\/th><td>0.1% Formic acid in water <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>0.1% Formic acid in acetonitrile <\/td><\/tr><tr><td><\/td><td><table class=\"cgram_ramp\"><thead><tr><th>Time (min)<\/th><th>Flow (mL\/min)<\/th><th>%A<\/th><th>%B<\/th><\/tr><\/thead><tbody><tr><td>0.00<\/td><td>0.6<\/td><td>5<\/td><td>95<\/td><\/tr><tr><td>2.00<\/td><td>0.6<\/td><td>70<\/td><td>30<\/td><\/tr><tr><td>2.01<\/td><td>0.6<\/td><td>5<\/td><td>95<\/td><\/tr><tr><td>3.50<\/td><td>0.6<\/td><td>5<\/td><td>95<\/td><\/tr><\/tbody><\/table><\/td><\/tr><\/table><table class=\"conditions col-lg-6 col-12\"><tr><th class=\"conditions_header\" scope=\"row\">Detector<\/th><td>MS\/MS<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Ion Mode:<\/th><td>ESI+ <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Mode:<\/th><td>MRM <\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Instrument<\/th><td>UHPLC<\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0559.pdf\" target=\"_blank\" rel=\"noopener noreferrer\"><svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"18\" height=\"18\" viewBox=\"0 0 18 18\"><g data-name=\"Group 2996\"><path data-name=\"Rectangle 1246\" d=\"M0 0h18v18H0z\" style=\"fill: none;\"><\/path><\/g><g data-name=\"Group 2997\"><path data-name=\"Path 729\" d=\"M13.412 11.4v2.017H5.345V11.4H4v2.017a1.349 1.349 0 0 0 1.345 1.345h8.068a1.349 1.349 0 0 0 1.345-1.345V11.4zm-.672-2.694-.948-.948-1.741 1.735V4H8.706v5.493L6.965 7.758l-.948.948 3.361 3.361z\" transform=\"translate(-.437 -.414)\" style=\"fill: rgb(13, 123, 196);\"><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><\/div><div class='chromatogram-article-inner-full'><div class=\"chromatogram-article-inner\">\n<div class=\"wp-block-custom-chromatogram-article\"><div class=\"wp-block-custom-chromatogram-article\"><div class=\"chromatogram-image regular-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0561.png\" alt=\"4-Methylimidazole Lifetime Over 500 Injection on Raptor FluoroPhenyl\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_FF0561<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12\">\n<thead><tr><th><\/th><th style=\"text-align: left;width: 75px\">Peaks<\/th><th style=\"text-align: center;width: 75px\">t<sub>R<\/sub> (min)<\/th><th style=\"text-align: center;width: 75px\">Precursor Ion<\/th><th style=\"text-align: center;width: 75px\">Product Ion<\/th><\/tr><\/thead>\n<tbody><tr><td class=\"num\">1.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 4-Methylimidazole (4-MEI)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/822-36-6\/4-Methylimidazole\" target=\"_blank\" rel=\"noopener\">4-Methylimidazole (4-MEI)<\/a><\/td><td class=\"oth\">1.19<\/td><td class=\"oth\">83<\/td><td class=\"oth\">56<\/td><\/tr>\n<\/tbody><\/table><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0561.pdf\" target=\"_blank\" rel=\"noopener noreferrer\"><svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"18\" height=\"18\" viewBox=\"0 0 18 18\"><g data-name=\"Group 2996\"><path data-name=\"Rectangle 1246\" d=\"M0 0h18v18H0z\" style=\"fill: none;\"><\/path><\/g><g data-name=\"Group 2997\"><path data-name=\"Path 729\" d=\"M13.412 11.4v2.017H5.345V11.4H4v2.017a1.349 1.349 0 0 0 1.345 1.345h8.068a1.349 1.349 0 0 0 1.345-1.345V11.4zm-.672-2.694-.948-.948-1.741 1.735V4H8.706v5.493L6.965 7.758l-.948.948 3.361 3.361z\" transform=\"translate(-.437 -.414)\" style=\"fill: rgb(13, 123, 196);\"><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Raptor FluoroPhenyl columns perform well in both HILIC and reversed phase modes, giving method developers the flexibility to select the mode that performs best for their specific compounds of interest. Here we use a Raptor Fluorophenyl column in HILIC mode for LC analysis of 4-methylimidazole (4-MEI). This approach provides more retention than reversed phase mode and can be applied to determining 4-MEI levels in caramel colorings to monitor formulations and also in foods and beverages to assess potential exposure.<\/p>\n","protected":false},"author":11,"featured_media":6901,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_kadence_starter_templates_imported_post":false,"_kad_post_transparent":"","_kad_post_title":"","_kad_post_layout":"","_kad_post_sidebar_id":"","_kad_post_content_style":"","_kad_post_vertical_padding":"","_kad_post_feature":"","_kad_post_feature_position":"","_kad_post_header":false,"_kad_post_footer":false,"footnotes":""},"categories":[789],"tags":[],"industries-application":[2274,2275],"post-badge":[],"resource-type":[2623],"product-library":[2571,2553],"resource-technique":[2357,2343,2345,2365],"hf_cat_post":[650,2624],"ppma_author":[414],"class_list":["post-43564","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-articles-ja","industries-application-food-beverage-ja","industries-application-food-safety-ja","resource-type-featured-application","product-library-lc-columns-ja","product-library-liquid-chromatography-products-ja","resource-technique-hilic-ja","resource-technique-liquid-chromatography-ja","resource-technique-mass-spectrometry-ms-ja","resource-technique-ms-ms-ja"],"acf":[],"taxonomy_info":{"category":[{"value":789,"label":"\u6280\u8853\u8cc7\u6599"}],"industries-application":[{"value":2274,"label":"\u98df\u54c1\uff5c\u98f2\u6599"},{"value":2275,"label":"\u98df\u306e\u5b89\u5168"}],"resource-type":[{"value":2623,"label":"Featured 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