{"id":43387,"date":"2020-10-13T00:00:00","date_gmt":"2020-10-13T00:00:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/fast-sensitive-lc-msms-analysis-of-ghb-and-related-compounds-in-human-blood-for-forensic-testing\/"},"modified":"2025-12-08T14:35:54","modified_gmt":"2025-12-08T14:35:54","slug":"fast-sensitive-lc-msms-analysis-of-ghb-and-related-compounds-in-human-blood-for-forensic-testing","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/de\/articles-ja\/cfss2892\/fast-sensitive-lc-msms-analysis-of-ghb-and-related-compounds-in-human-blood-for-forensic-testing","title":{"rendered":"Fast, Sensitive LC-MS\/MS Analysis of GHB and Related Compounds in Human Blood for Forensic Testing"},"content":{"rendered":"\n<ul class=\"wp-block-list\">\n<li>Simultaneously analyze GHB, GBL, and 1,4-BD in whole blood samples in a fast, 5-minute cycle time.<\/li>\n\n\n\n<li>Chromatographic separation of actual GBL and GBL produced from GHB in-source conversion.<\/li>\n\n\n\n<li>Sufficient sensitivity to measure endogenous levels of GHB and to identify exogenous drug ingestion.<\/li>\n<\/ul>\n\n\n\n<p>Recently, gamma-hydroxybutyrate (GHB) and its precursors gamma-butyrolactone (GBL) and 1,4-butanediol (1,4-BD) have been gaining popularity as recreational drugs because of their euphoric properties. In addition, due to its potent pro-sexual effects, GHB is increasingly implicated in drug-facilitated sexual assault. Because of their easy availability and potential for misuse, demand is growing for analytical determination of these drugs in both urine and blood samples. Analysis can be challenging because these compounds are small molecules that are difficult to retain, so many methods employ time-consuming derivatization and require long analysis times. Measurement is further complicated by both endogenous levels of GHB, which is present in all tissues as a naturally occurring neurotransmitter, and by the very rapid metabolism and elimination of ingested GHB.<\/p>\n\n\n\n<p>The LC-MS\/MS analysis of GHB and its precursors shown here uses a quick and simple protein crash procedure for whole blood samples. Excellent retention and chromatographic separations are reliably obtained in a fast, 5-minute analysis without the need for derivatization or extensive sample preparation. All compounds were chromatographically separated on a Force C18 column, which is critical for this analysis because GHB can convert to GBL in the instrument\u2019s ion source. In addition, method sensitivity is sufficient to measure low endogenous levels of GHB and to identify exogenous ingestion of these drugs. This LC-MS\/MS analysis of GHB and its precursors is a faster alternative to typical approaches found in the literature and provides an effective means of testing and reporting levels in biological specimens.<\/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 regular-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0732.png\" alt=\"GHB and Related Compounds in Human Blood by LC-MS\/MS\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0732<\/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\">\u03b3-Hydroxybutyric acid (GHB)<\/td><td class=\"oth\">1.42<\/td><td class=\"oth\">105.2<\/td><td class=\"oth\">87.0<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 1,4-Butanediol (1,4-BD)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/110-63-4\/1,4-Butanediol\" target=\"_blank\" rel=\"noopener\">1,4-Butanediol (1,4-BD)<\/a><\/td><td class=\"oth\">1.42<\/td><td class=\"oth\">91.0<\/td><td class=\"oth\">55.0<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for \u03b3-Butyrolactone (GBL)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/96-48-0\/\u03b3-Butyrolactone\" target=\"_blank\" rel=\"noopener\">\u03b3-Butyrolactone (GBL)<\/a><\/td><td class=\"oth\">1.75<\/td><td class=\"oth\">87.0<\/td><td class=\"oth\">45.0<\/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>Force C18  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/963431E?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0732\" rel=\"noopener\">cat.# 963431E<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Dimensions:<\/th><td>100 mm x 3.0 mm ID<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Particle Size:<\/th><td>3 \u00b5m<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Pore Size:<\/th><td>100 \u00c5<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Guard Column:<\/th><td>Force C18 EXP guard column cartridge 5 mm, 3.0 mm ID, 3 \u00b5m (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/963450253?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0732\" rel=\"noopener\">cat.# 963450253<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Temp.:<\/th><td>30 \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>Water<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>500 ng\/mL<\/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>0.5% Formic acid in water <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>0.5% Formic acid in methanol <\/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>0.7<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>0.50<\/td><td>0.7<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>3.00<\/td><td>0.7<\/td><td>50<\/td><td>50<\/td><\/tr><tr><td>3.01<\/td><td>0.7<\/td><td>95<\/td><td>5<\/td><\/tr><tr><td>5.00<\/td><td>0.7<\/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\">Sample Preparation<\/th><td>100 \u03bcL of whole human blood was fortified at 50 \u03bcg\/mL with GHB, GBL, 1,4-BD, and GHB-D6 (IS) using 5 \u03bcL of 1 mg\/mL solutions. The blood was precipitated with 380 \u03bcL methanol. The sample was then vortexed at 1000 rpm for 10 seconds and centrifuged at 3000 rpm for 10 minutes at 10 \u00b0C. 50 \u03bcL of the supernatant was removed and diluted to 1 mL with water. The sample was then vortexed and subjected to LC-MS\/MS analysis. (Internal standard not shown on chromatogram.)<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Notes<\/th><td>Want even better performance when analyzing metal-sensitive compounds? Check out Inert LC columns at <a href=\"https:\/\/www.restek.com\/inert\" target=\"_blank\" rel=\"noopener\">www.restek.com\/inert<\/a>.<\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0732.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>This straightforward method for LC-MS\/MS analysis of gamma-hydroxybutyrate (GHB) and its precursors gamma-butyrolactone and 1,4-butanediol provides simultaneous analysis of all target compounds in a fast, 5-min cycle time. Good low-level sensitivity was obtained, sufficient to measure low endogenous levels of GHB and identify exogenous drug consumption.<\/p>\n","protected":false},"author":11,"featured_media":6499,"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":[2276,2277],"post-badge":[],"resource-type":[2623],"product-library":[2571,2553],"resource-technique":[2343,2365],"hf_cat_post":[650,2624],"ppma_author":[414],"class_list":["post-43387","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-articles-ja","industries-application-forensics-ja","industries-application-forensics-toxicology-ja","resource-type-featured-application","product-library-lc-columns-ja","product-library-liquid-chromatography-products-ja","resource-technique-liquid-chromatography-ja","resource-technique-ms-ms-ja"],"acf":[],"taxonomy_info":{"category":[{"value":789,"label":"\u6280\u8853\u8cc7\u6599"}],"industries-application":[{"value":2276,"label":"\u6cd5\u533b\u5b66"},{"value":2277,"label":"\u6cd5\u4e2d\u6bd2\u5b66"}],"resource-type":[{"value":2623,"label":"Featured 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