{"id":43588,"date":"2020-10-28T00:00:00","date_gmt":"2020-10-28T00:00:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/rapid-simple-4-5-minute-pyrrolizidine-alkaloids-analysis-by-lc-msms\/"},"modified":"2026-01-28T16:59:25","modified_gmt":"2026-01-28T16:59:25","slug":"rapid-simple-4-5-minute-pyrrolizidine-alkaloids-analysis-by-lc-msms","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/es\/articles\/ffss3064\/rapid-simple-4-5-minute-pyrrolizidine-alkaloids-analysis-by-lc-msms","title":{"rendered":"Rapid, Simple 4.5-Minute Pyrrolizidine Alkaloids Analysis by LC-MS\/MS"},"content":{"rendered":"\n<ul class=\"wp-block-list\">\n<li>Fast 4.5-min pyrrolizidine alkaloids analysis separates most compounds to baseline.<\/li>\n\n\n\n<li>Simple mobile phase and gradient elutes compounds quickly with excellent peak shape.<\/li>\n\n\n\n<li>Robust Raptor ARC-18 column outlasts traditional C18 columns under acidic conditions.<\/li>\n<\/ul>\n\n\n\n<p>Pyrrolizidine alkaloids (PAs) are toxins found naturally in a wide variety of plant species (e.g., daisies, forget-me-nots, and legumes, among others). They are secondary metabolites formed to protect plants against herbivores and, according to the European Food Safety Authority (EFSA), are likely the most widely distributed natural toxins. While there are over 6000 plant species known to contain pyrrolizidine alkaloids, only a few species have been involved in the direct poisoning of people or animals. And, of the roughly 700 known PAs, the 1,2-unsaturated compounds have been identified as the most toxic, capable of causing fatal liver damage. Herbs, teas, honey, and cereal may be contaminated with pyrrolizidine alkaloids if plants containing PAs are accidentally co-harvested or used as a food source for bees.<\/p>\n\n\n\n<p>Because of the World Health Organization&#8217;s (WHO) Codex Alimentarius Commission adoption of the \u201cas low as reasonably achievable\u201d (ALARA) principle for PAs\u2014along with many regional efforts to monitor these toxic compounds in food, feed, and medicines, the need for fast, accurate pyrrolizidine alkaloids analysis methods is on the rise. At present, the only harmonized method was developed and validated by the German Federal Institute for Risk Assessment (BfR) [1]. It employs a sulfuric acid digestion, an SPE C18 cleanup, and analysis via LC-MS\/MS. The example method provided in the BfR protocol uses a 150 x 2.1 mm, 1.9 \u00b5m fully porous particle C18 column and produces a run time of 15 minutes with the last compound, lasiocarpine-N-oxide (LaN), eluting at 9.33 minutes. However, in the case study presented below, switching to a superficially porous 50 x 2.1 mm, 2.7 \u00b5m Raptor ARC-18 column provides sufficient retention while still eluting LaN in just 3.2 minutes (4.5 minute total run time). Note that although a UHPLC instrument was used in this example, it is not necessary for this analysis. Excellent peak shapes were obtained and all compounds, except for echimidine and its corresponding N-oxide, were separated to baseline. The Raptor ARC-18 column\u2019s unique phase chemistry also results in longer lifetimes under acidic conditions compared to traditional C18 columns.<\/p>\n\n\n\n<p>As the scope of pyrrolizidine alkaloids analysis continues to expand, methods will have to grow along with it, but for today\u2019s challenges, using a Raptor ARC-18 column allows labs to optimize method conditions, shorten the overall run time, and increase sample throughput.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">References<\/h2>\n\n\n\n<ol class=\"wp-block-list\">\n<li>Bundesinstitut f\u00fcr Risikobewertung (BfR), BfR-PA-Tee-2.0\/2014, Bestimmung von Pyrrolizidinalkaloiden (PA) in Pflanzenmaterial mittels SPE-LC-MS\/MS, 2014.&nbsp;<a href=\"https:\/\/www.bfr.bund.de\/cm\/343\/bestimmung-von-pyrrolizidinalkaloiden.pdf\" target=\"_blank\" rel=\"noopener\">https:\/\/www.bfr.bund.de\/cm\/343\/bestimmung-von-pyrrolizidinalkaloiden.pdf<\/a><\/li>\n<\/ol>\n\n\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_ff0573.png\" alt=\"Pyrrolizidine Alkaloids on Raptor ARC-18 by LC-MS\/MS\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_FF0573<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks 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<\/th><th style=\"text-align: center;width: 75px\">Product 1<\/th><th style=\"text-align: center;width: 75px\">Product 2<\/th><th style=\"text-align: center;width: 75px\">Product 3<\/th><\/tr><\/thead>\n<tbody><tr><td class=\"num\">1.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Monocrotaline\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/315-22-0\/Monocrotaline\" target=\"_blank\" rel=\"noopener\">Monocrotaline<\/a><\/td><td class=\"oth\">0.677<\/td><td class=\"oth\">326.20<\/td><td class=\"oth\">120.3<\/td><td class=\"oth\">121.30<\/td><td class=\"oth\">94.25<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\">Incidine<\/td><td class=\"oth\">1.111<\/td><td class=\"oth\">300.20<\/td><td class=\"oth\">94.15<\/td><td class=\"oth\">138.20<\/td><td class=\"oth\">156.36<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Heliotrine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/303-33-3\/Heliotrine\" target=\"_blank\" rel=\"noopener\">Heliotrine<\/a><\/td><td class=\"oth\">1.763<\/td><td class=\"oth\">314.30<\/td><td class=\"oth\">138.20<\/td><td class=\"oth\">156.20<\/td><td class=\"oth\">120.20<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\">Heliotrine N-oxide<\/td><td class=\"oth\">1.895<\/td><td class=\"oth\">330.25<\/td><td class=\"oth\">172.25<\/td><td class=\"oth\">138.20<\/td><td class=\"oth\"><\/td><\/tr>\n<tr><td class=\"num\">5.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Senecionine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/130-01-8\/Senecionine\" target=\"_blank\" rel=\"noopener\">Senecionine<\/a><\/td><td class=\"oth\">2.240<\/td><td class=\"oth\">336.20<\/td><td class=\"oth\">120.30<\/td><td class=\"oth\">138.30<\/td><td class=\"oth\"><\/td><\/tr>\n<tr><td class=\"num\">6.<\/td><td class=\"cmpd\">Senecionine N-oxide<\/td><td class=\"oth\">2.410<\/td><td class=\"oth\">352.35<\/td><td class=\"oth\">94.20<\/td><td class=\"oth\">120.30<\/td><td class=\"oth\">118.25<\/td><\/tr>\n<tr><td class=\"num\">7.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Echimidine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/520-68-3\/Echimidine\" target=\"_blank\" rel=\"noopener\">Echimidine<\/a><\/td><td class=\"oth\">2.524<\/td><td class=\"oth\">398.30<\/td><td class=\"oth\">120.25<\/td><td class=\"oth\">220.15<\/td><td class=\"oth\"><\/td><\/tr>\n<tr><td class=\"num\">8.<\/td><td class=\"cmpd\">Echimidine N-oxide<\/td><td class=\"oth\">2.528<\/td><td class=\"oth\">414.25<\/td><td class=\"oth\">396.30<\/td><td class=\"oth\">352.20<\/td><td class=\"oth\">254.25<\/td><\/tr>\n<tr><td class=\"num\">9.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Lasiocarpine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/303-34-4\/Lasiocarpine\" target=\"_blank\" rel=\"noopener\">Lasiocarpine<\/a><\/td><td class=\"oth\">3.028<\/td><td class=\"oth\">412.20<\/td><td class=\"oth\">120.20<\/td><td class=\"oth\">220.15<\/td><td class=\"oth\">336.25<\/td><\/tr>\n<tr><td class=\"num\">10.<\/td><td class=\"cmpd\">Lasiocarpine N-oxide<\/td><td class=\"oth\">3.260<\/td><td class=\"oth\">428.25<\/td><td class=\"oth\">254.25<\/td><td class=\"oth\">352.25<\/td><td class=\"oth\">120.30<\/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 ARC-18  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9314A52?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_FF0573\" rel=\"noopener\">cat.# 9314A52<\/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><th class=\"sub conditions_header\" scope=\"row\">Pore Size:<\/th><td>90 \u00c5<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Temp.:<\/th><td>25 \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>0.1% Formic acid in water<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>10 ng\/mL<\/td><\/tr><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Inj. Vol.:<\/th><td>1 \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>0.5<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>3.5<\/td><td>0.5<\/td><td>65<\/td><td>35<\/td><\/tr><tr><td>3.51<\/td><td>0.5<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>4.5<\/td><td>0.5<\/td><td>95<\/td><td>5<\/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 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_ff0573.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>","protected":false},"excerpt":{"rendered":"<p>Pyrrolizidine alkaloid analysis is time consuming, with a typical run time of 15 minutes, and the acid digestion step can shorten analytical column lifetime. In contrast, this analysis using an acid-resistant Raptor ARC-18 column eluted the final compound in just 3.2 minutes with good peak shape and separation.<\/p>\n","protected":false},"author":11,"featured_media":6937,"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":[7],"tags":[],"industries-application":[2167,2169],"post-badge":[],"resource-type":[2623],"product-library":[2391,2373],"resource-technique":[2299,2301,2302],"hf_cat_post":[650,2624],"ppma_author":[414],"class_list":["post-43588","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-articles","industries-application-food-beverage","industries-application-food-safety","resource-type-featured-application","product-library-lc-columns","product-library-liquid-chromatography-products","resource-technique-liquid-chromatography","resource-technique-mass-spectrometry-ms","resource-technique-ms-ms"],"acf":[],"taxonomy_info":{"category":[{"value":7,"label":"Articles"}],"industries-application":[{"value":2167,"label":"Food &amp; 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