{"id":43385,"date":"2020-10-13T01:00:00","date_gmt":"2020-10-13T01:00:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/japanese-lc-msms-analysis-of-urinary-free-cortisol-and-cortisone-without-matrix-interferences-raptor-biphenyl-column-provides-reliable-separation-of-analytes-from-endogenous-matrix-interferences\/"},"modified":"2025-12-03T17:37:25","modified_gmt":"2025-12-03T17:37:25","slug":"japanese-lc-msms-analysis-of-urinary-free-cortisol-and-cortisone-without-matrix-interferences-raptor-biphenyl-column-provides-reliable-separation-of-analytes-from-endogenous-matrix-interferences","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/ja\/articles-ja\/cfss2824-ja\/japanese-lc-msms-analysis-of-urinary-free-cortisol-and-cortisone-without-matrix-interferences-raptor-biphenyl-column-provides-reliable-separation-of-analytes-from-endogenous-matrix-interferences","title":{"rendered":"\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u5e72\u6e09\u306e\u306a\u3044\u5c3f\u4e2d\u904a\u96e2\u30b3\u30eb\u30c1\u30be\u30ed\u30fc\u30eb\u3068\u30b3\u30eb\u30c1\u30be\u30f3\u306eLC-MS\/MS\u5206\u6790"},"content":{"rendered":"\n<ul class=\"wp-block-list\">\n<li>Raptor Biphenyl\u306f\u5185\u56e0\u6027\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u5e72\u6e09\u304b\u3089\u5206\u6790\u5bfe\u8c61\u5316\u5408\u7269\u306e\u78ba\u5b9f\u306a\u5206\u96e2\u3092\u53ef\u80fd\u306b\u3057\u307e\u3059\u3002<\/li>\n\n\n\n<li>C18\u3084\u4ed6\u306e\u30d5\u30a7\u30cb\u30eb\u7cfb\u56fa\u5b9a\u76f8\u3067\u306f\u5206\u96e2\u304c\u56f0\u96e3\u306a\u5316\u5408\u7269\u306e\u9078\u629e\u6027\u304c\u9ad8\u307e\u308a\u3001\u6b63\u78ba\u306a\u7d50\u679c\u304c\u5f97\u3089\u308c\u307e\u3059\u3002<\/li>\n\n\n\n<li>3\u5206\u306e\u9ad8\u901f\u5206\u6790\u306b\u3088\u3063\u3066\u3055\u3089\u306a\u308b\u30b5\u30f3\u30d7\u30eb\u306e\u30b9\u30eb\u30fc\u30d7\u30c3\u30c8\u306e\u5411\u4e0a\u304c\u53ef\u80fd\u3068\u306a\u308a\u307e\u3059\u3002<\/li>\n<\/ul>\n\n\n\n<p>\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u306f\u3001\u526f\u814e\u306b\u3066\u591a\u9175\u7d20\u306b\u3088\u308b\u30ab\u30b9\u30b1\u30fc\u30c9\u53cd\u5fdc\u306b\u3088\u3063\u3066\u30b3\u30ec\u30b9\u30c6\u30ed\u30fc\u30eb\u304b\u3089\u5408\u6210\u3055\u308c\u308b\u30b9\u30c6\u30ed\u30a4\u30c9\u30db\u30eb\u30e2\u30f3\u3067\u3059\u3002\u305d\u308c\u306f\u30d2\u30c8\u306b\u304a\u3051\u308b\u4e3b\u8981\u306a\u30b0\u30eb\u30b3\u30b3\u30eb\u30c1\u30b3\u30a4\u30c9\u3067\u3042\u308a\u3001\u305d\u3057\u3066\u30b0\u30eb\u30b3\u30fc\u30b9\u4ee3\u8b1d\u304a\u3088\u3073\u514d\u75ab\u5fdc\u7b54\u306e\u5236\u5fa1\u306b\u304a\u3044\u3066\u91cd\u8981\u306a\u5f79\u5272\u3092\u679c\u305f\u3057\u3066\u3044\u307e\u3059\u3002\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u306e\u4e0b\u6d41\u4ee3\u8b1d\u7523\u7269\u3067\u3042\u308b\u30b3\u30eb\u30c1\u30be\u30f3\u306f\u3001\u3055\u307e\u3056\u307e\u306a\u526f\u814e\u76ae\u8cea\u969c\u5bb3\u3001\u7279\u306b\u526f\u814e\u76ae\u8cea\u6a5f\u80fd\u4ea2\u9032\u75c7\uff08\u30af\u30c3\u30b7\u30f3\u30b0\u75c7\u5019\u7fa4\uff09\u306e\u8a3a\u65ad\u306b\u5f79\u7acb\u3064\u3088\u3046\u306b\u3001\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u3068\u540c\u6642\u306b\u6e2c\u5b9a\u3055\u308c\u307e\u3059\u3002\u5c3f\u4e2d\u904a\u96e2\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u304a\u3088\u3073\u30b3\u30eb\u30c1\u30be\u30f3\u30ec\u30d9\u30eb\u306f\u751f\u7406\u6d3b\u6027\u3092\u793a\u3059\u8840\u6f3f\u4e2d\u306e\u6fc3\u5ea6\u3068\u826f\u3044\u76f8\u95a2\u304c\u3042\u308a\u3001\u91cd\u8981\u306a\u30d0\u30a4\u30aa\u30de\u30fc\u30ab\u30fc\u3067\u3059\u3002LC-MS \/ MS\u306f\u9078\u629e\u6027\u304c\u9ad8\u304f\u3001\u30a4\u30e0\u30ce\u30a2\u30c3\u30bb\u30a4\u306b\u57fa\u3065\u304f\u65b9\u6cd5\u306b\u5f71\u97ff\u3092\u4e0e\u3048\u308b\u53ef\u80fd\u6027\u306e\u3042\u308b\u3001\u5206\u6790\u306b\u304a\u3051\u308b\u5e72\u6e09\u3092\u6392\u9664\u3059\u308b\u306e\u306b\u5f79\u7acb\u3064\u305f\u3081\u3001\u3053\u308c\u3089\u306e\u5316\u5408\u7269\u306b\u9069\u3057\u305f\u30b9\u30af\u30ea\u30fc\u30cb\u30f3\u30b0\u6280\u8853\u3067\u3059\u3002<\/p>\n\n\n\n<p>\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u3068\u30b3\u30eb\u30c1\u30be\u30f3\u306e\u69cb\u9020\u7684\u306a\u985e\u4f3c\u6027\u3001\u304a\u3088\u3073\u5c3f\u6210\u5206\u304b\u3089\u306e\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u52b9\u679c\u306e\u53ef\u80fd\u6027\u306b\u3088\u308a\u3001\u6b63\u78ba\u306a\u7d50\u679c\u3092\u5f97\u308b\u305f\u3081\u306b\u306f\u9069\u5207\u306a\u30ab\u30e9\u30e0\u9078\u629e\u6027\u304c\u91cd\u8981\u3067\u3059\u3002\u3053\u3053\u3067\u958b\u767a\u3055\u308c\u305f\u5c3f\u4e2d\u904a\u96e2\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u3068\u30b3\u30eb\u30c1\u30be\u30f3\u306eLC-MS\/MS\u5206\u6790\u3067\u306f\u3001\u9ad8\u3044\u9078\u629e\u6027\u3092\u6709\u3059\u308bRaptor Biphenyl\u30ab\u30e9\u30e0\u3092\u4f7f\u7528\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u4e21\u65b9\u306e\u5316\u5408\u7269\u304c\u5185\u56e0\u6027\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u5e72\u6e09\u304b\u3089\u5b8c\u5168\u306b\u5206\u96e2\u3055\u308c\u307e\u3057\u305f\u3002\u3044\u304f\u3064\u304b\u306e\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u5316\u5408\u7269\u306f\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u3068\u30a2\u30a4\u30bd\u30d0\u30ea\u30c3\u30af\u3067\u3042\u308a\u3001\u8cea\u91cf\u5206\u6790\u8a08\u306b\u3088\u3063\u3066\u5224\u5225\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u306a\u3044\u306e\u3067\u3001\u3053\u306e\u5834\u5408\u306f\u30af\u30ed\u30de\u30c8\u30b0\u30e9\u30d5\u30a3\u30fc\u5206\u96e2\u304c\u4e0d\u53ef\u6b20\u3067\u3057\u305f\u3002Raptor Biphenyl\u30ab\u30e9\u30e0\u3068\u3053\u3053\u306b\u793a\u3059\u6761\u4ef6\u3092\u4f7f\u7528\u3057\u3066\u30013\u5206\u306e\u5206\u6790\u3067\u30d9\u30fc\u30b9\u30e9\u30a4\u30f3\u5206\u96e2\u304c\u5f97\u3089\u308c\u307e\u3057\u305f\u3002\u3053\u308c\u306b\u3088\u308a\u3001\u81e8\u5e8a\u8a3a\u65ad\u306e\u305f\u3081\u306e\u3088\u308a\u6b63\u78ba\u306a\u5b9a\u91cf\u30c7\u30fc\u30bf\u3092\u8fc5\u901f\u306b\u751f\u6210\u3067\u304d\u307e\u3059\u3002<\/p>\n\n\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_cf0697.png\" alt=\"Cortisol and Cortisone in Extracted Female Human Urine on Raptor Biphenyl by LC-MS\/MS\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0697<\/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><th style=\"text-align: center;width: 75px\">Product Ion<\/th><\/tr><\/thead>\n<tbody><tr><td class=\"num\">1.<\/td><td class=\"cmpd\">Cortisol-d4<\/td><td class=\"oth\">0.75<\/td><td class=\"oth\">367.4<\/td><td class=\"oth\">121.0<\/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 Endogenous cortisol\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/50-23-7\/Endogenous cortisol\" target=\"_blank\" rel=\"noopener\">Endogenous cortisol<\/a><\/td><td class=\"oth\">0.76<\/td><td class=\"oth\">363.3<\/td><td class=\"oth\">120.9<\/td><td class=\"oth\">91.1<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\">Cortisone-d8<\/td><td class=\"oth\">0.88<\/td><td class=\"oth\">369.4<\/td><td class=\"oth\">167.9<\/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 Endogenous cortisone\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/53-06-5\/Endogenous cortisone\" target=\"_blank\" rel=\"noopener\">Endogenous cortisone<\/a><\/td><td class=\"oth\">0.89<\/td><td class=\"oth\">361.3<\/td><td class=\"oth\">163.2<\/td><td class=\"oth\">91.0<\/td><\/tr>\n<\/tbody><\/table><div class=\"peaknotes\">*Isobaric matrix peaks<\/div><\/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 Biphenyl  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9309A52?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0697\" rel=\"noopener\">cat.# 9309A52<\/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\">Guard Column:<\/th><td>Raptor Biphenyl EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 \u00b5m (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9309A0252?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0697\" rel=\"noopener\">cat.# 9309A0252<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Temp.:<\/th><td>40 \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>Mobile phase A<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>  Calculated concentration is 105.1 ng\/mL and 135.9 ng\/mL for cortisol and cortisone, respectively<\/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>Water, 0.1% formic acid <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>Acetonitrile, 0.1% formic acid <\/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>70<\/td><td>30<\/td><\/tr><tr><td>1.00<\/td><td>0.6<\/td><td>70<\/td><td>30<\/td><\/tr><tr><td>1.01<\/td><td>0.6<\/td><td>0<\/td><td>100<\/td><\/tr><tr><td>1.50<\/td><td>0.6<\/td><td>0<\/td><td>100<\/td><\/tr><tr><td>1.51<\/td><td>0.6<\/td><td>70<\/td><td>30<\/td><\/tr><tr><td>3.00<\/td><td>0.6<\/td><td>70<\/td><td>30<\/td><\/tr><\/tbody><\/table><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Max Pressure:<\/th><td>400 bar<\/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><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Sample Preparation<\/th><td>1. Centrifuge female human urine for 5 min at 4500 rpm, 10 \u00b0C.<br \/>2. Aliquot 380 \u00b5L supernatant. Add 20 \u00b5L each of internal standard solution (1 \u00b5g\/mL in methanol). <br \/>3. Load 200 \u03bcL of sample on to ISOLUTE SLE+ 200 \u00b5L supported liquid extraction plate (part# 820-0200-P01). <br \/>4. Apply a pulse of vacuum to initiate flow.<br \/>5. Wait 5 min for sample to completely absorb.<br \/>6. Extract samples with 1 mL of MTBE. Allow solvent to flow for 5 min under gravity. Apply vacuum for 10-30 sec to complete elution.<br \/>7. Evaporate extracts to dryness under a stream of nitrogen.<br \/>8. Reconstitute in 200 \u00b5L mobile phase A prior to analysis.<br \/>9. Vortex to mix.<br \/><\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0697.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\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n        <div class=\"cpb\">\n            <h3 class=\"cpb-heading\">Products Mentioned<\/h3>\n            <hr class=\"cpb-heading-underline\" \/>\n            <div class=\"cpb-list\">\n                                    <div class=\"cpb-item\">\n                        <div class=\"cpb-col cpb-col--left\">\n                            <a class=\"cpb-catalog\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309A52\">                                Catalog No. 9309A52                            <\/a>                        <\/div>\n                        <div class=\"cpb-col cpb-col--middle\">\n                            <div class=\"cpb-title\">Raptor Biphenyl, 2.7 \u00b5m, 50 x 2.1 mm HPLC\u30ab\u30e9\u30e0<\/div>\n                        <\/div>\n                        <div class=\"cpb-col cpb-col--right\">\n                                                            <a class=\"cpb-view-btn\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309A52\">\u88fd\u54c1\u60c5\u5831\u3092\u898b\u308b<\/a>\n                                                    <\/div>\n                    <\/div>\n                                    <div class=\"cpb-item\">\n                        <div class=\"cpb-col cpb-col--left\">\n                            <a class=\"cpb-catalog\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309A0252\">                                Catalog No. 9309A0252                            <\/a>                        <\/div>\n                        <div class=\"cpb-col cpb-col--middle\">\n                            <div class=\"cpb-title\">Raptor Biphenyl EXP Guard Column Cartridge, 2.7 \u00b5m, 5 x 2.1 mm, 3-pk.<\/div>\n                        <\/div>\n                        <div class=\"cpb-col cpb-col--right\">\n                                                            <a class=\"cpb-view-btn\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309A0252\">\u88fd\u54c1\u60c5\u5831\u3092\u898b\u308b<\/a>\n                                                    <\/div>\n                    <\/div>\n                            <\/div>\n        <\/div>\n        \n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>\u4e00\u822c\u306b\u3001LC-MS\/MS\u306b\u3088\u308b\u5c3f\u4e2d\u306e\u904a\u96e2\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u304a\u3088\u3073\u30b3\u30eb\u30c1\u30be\u30f3\u5206\u6790\u306f\u3001\u30a4\u30e0\u30ce\u30a2\u30c3\u30bb\u30a4\u306b\u57fa\u3065\u304f\u65b9\u6cd5\u3088\u308a\u3082\u512a\u308c\u305f\u7279\u7570\u6027\u3092\u3082\u305f\u3089\u3057\u307e\u3059\u3002\u3057\u304b\u3057\u3001\u30b3\u30eb\u30c1\u30be\u30fc\u30eb\u3092\u30a2\u30a4\u30bd\u30d0\u30ea\u30c3\u30af\u5316\u5408\u7269\u304b\u3089\u5206\u96e2\u3059\u308b\u9078\u629e\u6027\u304c\u30ab\u30e9\u30e0\u306b\u306f\u6c42\u3081\u3089\u308c\u307e\u3059\u3002\u3053\u3053\u3067\u7d39\u4ecb\u3059\u308b\u65b9\u6cd5\u3067\u306f\u3001Raptor 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