{"id":43364,"date":"2020-10-12T00:00:00","date_gmt":"2020-10-12T00:00:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/tech-tip-column-conditioning-ensures-consistent-etgets-results\/"},"modified":"2025-12-10T14:42:33","modified_gmt":"2025-12-10T14:42:33","slug":"tech-tip-column-conditioning-ensures-consistent-etgets-results","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/zh-hans\/articoli\/cfar2737\/tech-tip-column-conditioning-ensures-consistent-etgets-results","title":{"rendered":"Tech Tip: Column Conditioning Ensures Consistent EtG\/EtS Results"},"content":{"rendered":"\n<p>Typical methods for analyzing EtG and EtS in human urine have several limitations: poor retention and resolution of EtG and EtS from matrix components, long run times that limit sample throughput, and short column lifetimes. Restek\u2019s chemists have developed a simple, dilute-and-shoot LC-MS\/MS method for EtG\/EtS analysis using the Raptor EtG\/EtS column, which provides consistent separations in a fast, four-minute analysis time.<\/p>\n\n\n\n<p>The Raptor EtG\/EtS column features a novel stationary phase developed specifically for this critical application. This method was developed and stringently tested on the Raptor EtG\/EtS column because it provides the retention needed to consistently elute the target analytes well away from matrix interferences and is rugged and long-lasting to satisfy the demands of high-throughput labs. As with all new columns, it is good practice to run a number of conditioning or equilibration injections in order to ensure good response, peak shape, and retention time consistency. As part of method development, we determined that running 30 matrix injections through the full gradient program on a new column ensured highly consistent performance during subsequent sample analysis (Figure 1).<\/p>\n\n\n\n<p>By properly conditioning new columns and employing Restek\u2019s EtG\/EtS method for sample prep and analysis, high-throughput labs testing human urine samples for these alcohol consumption biomarkers can ensure accurate, consistent results with fewer column changes. Full method parameters, verification data, and analyses of patient samples are provided&nbsp;<a href=\"https:\/\/discover.restek.com\/cfar3061\/successful-strategies-for-the-analysis-of-etg-and-ets-in-urine-01\/\" data-type=\"post\" data-id=\"43365\">here<\/a>.<\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 1:<\/strong>\u00a0Highly consistent EtG\/EtS results are obtained using a preconditioned Raptor EtG\/EtS column and the method parameters shown here. Matrix interferences are still well resolved even after 1,000 sample injections.<\/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_cf0706.png\" alt=\"1,000 Injection Lifetime: EtG and EtS in Urine on Raptor EtG\/EtS\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0706<\/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\">Precursor Ion<\/th><th style=\"text-align: center;width: 75px\">Product 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 Ethyl-\u03b2-D-glucuronide-d5\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/1135070-98-2\/Ethyl-\u03b2-D-glucuronide-d5\" target=\"_blank\" rel=\"noopener\">Ethyl-\u03b2-D-glucuronide-d5<\/a><\/td><td class=\"oth\">225.9<\/td><td class=\"oth\">84.9<\/td><td class=\"oth\">&#8211;<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Ethyl-\u03b2-D-glucuronide\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/17685-04-0\/Ethyl-\u03b2-D-glucuronide\" target=\"_blank\" rel=\"noopener\">Ethyl-\u03b2-D-glucuronide<\/a><\/td><td class=\"oth\">220.8<\/td><td class=\"oth\">84.9<\/td><td class=\"oth\">74.8<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Ethyl sulfate-d5\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/1329611-05-3\/Ethyl sulfate-d5\" target=\"_blank\" rel=\"noopener\">Ethyl sulfate-d5<\/a><\/td><td class=\"oth\">129.7<\/td><td class=\"oth\">97.7<\/td><td class=\"oth\">&#8211;<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Ethyl sulfate\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/64-67-5\/Ethyl sulfate\" target=\"_blank\" rel=\"noopener\">Ethyl sulfate<\/a><\/td><td class=\"oth\">124.7<\/td><td class=\"oth\">96.8<\/td><td class=\"oth\">79.7<\/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 EtG\/EtS  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9325A12?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 9325A12<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Dimensions:<\/th><td>100 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\">Guard Column:<\/th><td>UltraShield UHPLC precolumn filter, 0.2 \u00b5m frit (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/25809?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 25809<\/a>)<\/td><\/tr><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>0.1% Formic acid in water<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>  A 500 ng\/mL QC sample was prepared in urine. 50 \u00b5L of the sample was diluted with 950 \u00b5L of a working internal standard (25 ng\/mL EtS-d5\/100 ng\/mL EtG-d5 in 0.1% formic acid in water). The sample was vortexed at 3500 rpm for 10 seconds to mix. The sample was then centrifuged at 3000 rpm for 5 minutes at 10 \u00b0C. The autosampler needle was adjusted to inject from the supernatant.<\/td><\/tr><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Inj. Vol.:<\/th><td>10 \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>2.50<\/td><td>0.5<\/td><td>65<\/td><td>35<\/td><\/tr><tr><td>2.51<\/td><td>0.5<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>4.00<\/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><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>HPLC<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Notes<\/th><td><b>Reference Standards<\/b><br \/>Ethyl-\u03b2-D-glucuronide (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/34101?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 34101<\/a>)<br \/>Ethyl-\u03b2-D-glucuronide-d5 (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/34102?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 34102<\/a>)<br \/>Ethyl sulfate sodium salt (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/34103?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 34103<\/a>)<br \/>Ethyl sulfate-d5 sodium salt (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/34104?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0706\" rel=\"noopener\">cat.# 34104<\/a>)<br \/><\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0706.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<p><\/p>\n\n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Methods for monitoring alcohol consumption biomarkers EtG and EtS are generally limited by poor retention and coelution with matrix interferences, as well as by long analysis times and short column lifetimes. The dilute-and-shoot EtG\/EtS LC-MS\/MS analysis developed here using the novel Raptor EtG\/EtS column easily resolves EtG and EtS from matrix interferences, providing consistent, accurate results for high-throughput labs testing human urine samples for alcohol consumption.<\/p>\n","protected":false},"author":11,"featured_media":6439,"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":[788],"tags":[],"industries-application":[2245,2248,2260,2261],"post-badge":[],"resource-type":[563],"product-library":[2535,2517],"resource-technique":[2334,2336,2364],"hf_cat_post":[650],"ppma_author":[414],"class_list":["post-43364","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-articoli","industries-application-clinico","industries-application-tossicologia-clinica","industries-application-forense","industries-application-tossicologia-forense","resource-type-column-installation-and-care","product-library-colonne-lc","product-library-prodotti-per-cromatografia-liquida","resource-technique-cromatografia-liquida-lc","resource-technique-spettometria-di-massa-ms","resource-technique-ms-ms-it"],"acf":[],"taxonomy_info":{"category":[{"value":788,"label":"Articoli"}],"industries-application":[{"value":2245,"label":"Clinico"},{"value":2248,"label":"Tossicologia 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