{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T19:45:05Z","timestamp":1787168705361,"version":"build-2736575974"},"reference-count":18,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2024,6,19]],"date-time":"2024-06-19T00:00:00Z","timestamp":1718755200000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2024,6]]},"abstract":"Abstract<\/jats:title>\n \n Fabry disease (FD) is a lysosomal storage disorder caused by variants in the\n GLA<\/jats:italic>\n gene encoding \u03b1\u2010galactosidase A, an enzyme required for catabolism of globotriaosylceramide (Gb\n 3<\/jats:sub>\n ). Accumulation of Gb\n 3<\/jats:sub>\n in patients\u2019 cells, tissues, and biological fluids causes clinical manifestations including ventricular hypertrophy, renal insufficiency, and strokes. This protocol describes a methodology to analyze urinary Gb\n 3<\/jats:sub>\n and creatinine. Samples are diluted with an internal standard solution containing Gb\n 3<\/jats:sub>\n (C17:0) and creatinine\u2010D\n 3<\/jats:sub>\n , centrifuged, and directly analyzed by ultra\u2010high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC\u2010MS\/MS) using an 8.7\u2010min method. Eight Gb\n 3<\/jats:sub>\n isoforms [C16:0, C18:0, C20:0, C22:1, C22:0, C24:1, C24:0, and (C24:0)OH] are analyzed and the total is normalized to creatinine. Confirmation ions are monitored to detect potential interferences. The Gb\n 3<\/jats:sub>\n limit of quantification is 0.023 \u00b5g\/ml. Its interday coefficients of variation (3 concentrations measured) are \u226415.4%. This method minimizes matrix effects (\u22646.5%) and prevents adsorption or precipitation of Gb\n 3<\/jats:sub>\n . Urine samples are stable (bias <15%) for 2 days at 21\u00b0C, 7 days at 4\u00b0C, and 4 freeze\/thaw cycles, whereas prepared samples are stable for 5 days at 21\u00b0C, and 14 days at 4\u00b0C. The Gb\n 3<\/jats:sub>\n \/creatinine age\u2010related upper reference limits (mean + 2 standard deviations) are 29 mg\/mol creatinine (<7 years) and 14 mg\/mol creatinine (\u22657 years). This simple, robust protocol has been fully validated (ISO 15189) and provides a valuable tool for diagnosis and monitoring of FD patients. \u00a9 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.\n <\/jats:p>\n \n Basic Protocol<\/jats:bold>\n : Analysis of urinary globotriaosylceramide (Gb\n 3<\/jats:sub>\n ) and creatinine by UHPLC\u2010MS\/MS\n <\/jats:p>\n \n Support Protocol 1<\/jats:bold>\n : Preparation of the urinary quality controls\n <\/jats:p>\n \n Support Protocol 2<\/jats:bold>\n : Preparation of the urine matrix used for the Gb\n 3<\/jats:sub>\n calibration curve\n <\/jats:p>\n \n Support Protocol 3<\/jats:bold>\n : Preparation of the Gb\n 3<\/jats:sub>\n calibrators\n <\/jats:p>\n \n Support Protocol 4<\/jats:bold>\n : Preparation of the working solution containing the internal standards\n <\/jats:p>\n \n Support Protocol 5<\/jats:bold>\n : Preparation of the creatinine calibrators\n <\/jats:p>\n \n Support Protocol 6<\/jats:bold>\n : Preparation of the UHPLC solutions and mobile phases\n <\/jats:p>","DOI":"10.1002\/cpz1.1087","type":"journal-article","created":{"date-parts":[[2024,6,19]],"date-time":"2024-06-19T10:23:22Z","timestamp":1718792602000},"source":"Crossref","is-referenced-by-count":1,"title":["Analysis of Globotriaosylceramide (Gb\n 3<\/sub>\n ) in Liquid Urine: A Straightforward Assay Using Tandem Mass Spectrometry"],"prefix":"10.1002","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6289-1839","authenticated-orcid":false,"given":"Michel","family":"Boutin","sequence":"first","affiliation":[{"name":"Division of Medical Genetics, Department of Laboratory Medicine CIUSSS de l'Estrie\u2010CHUS Sherbrooke Quebec Canada"},{"name":"Department of Pediatrics Universit\u00e9 de Sherbrooke Sherbrooke Quebec Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6685-600X","authenticated-orcid":false,"given":"Bruno","family":"Maranda","sequence":"additional","affiliation":[{"name":"Division of Medical Genetics, Department of Laboratory Medicine CIUSSS de l'Estrie\u2010CHUS Sherbrooke Quebec Canada"},{"name":"Department of Pediatrics Universit\u00e9 de Sherbrooke Sherbrooke Quebec Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4898-1859","authenticated-orcid":false,"given":"Paula J.","family":"Waters","sequence":"additional","affiliation":[{"name":"Division of Medical Genetics, Department of Laboratory Medicine CIUSSS de l'Estrie\u2010CHUS Sherbrooke Quebec Canada"},{"name":"Department of Pediatrics Universit\u00e9 de Sherbrooke Sherbrooke Quebec Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,6,19]]},"reference":[{"key":"e_1_2_12_2_1_1","doi-asserted-by":"publisher","DOI":"10.1002\/cphg.24"},{"key":"e_1_2_12_2_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/cphg.34"},{"key":"e_1_2_12_2_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.7b03609"},{"key":"e_1_2_12_2_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21176114"},{"key":"e_1_2_12_2_5_1","doi-asserted-by":"publisher","DOI":"10.3390\/jcm1008166"},{"key":"e_1_2_12_2_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40620\u2010015\u20100193\u20101"},{"key":"e_1_2_12_2_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbadis.2019.07.005"},{"key":"e_1_2_12_2_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jfma.2023.10.004"},{"key":"e_1_2_12_2_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/genes15010037"},{"key":"e_1_2_12_2_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13023\u2010024\u201003028\u2010w"},{"key":"e_1_2_12_2_11_1","doi-asserted-by":"publisher","DOI":"10.1159\/000448968"},{"key":"e_1_2_12_2_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2018.02.014"},{"key":"e_1_2_12_2_13_1","doi-asserted-by":"publisher","DOI":"10.2174\/1570159x20666220601124117"},{"key":"e_1_2_12_2_14_1","doi-asserted-by":"publisher","DOI":"10.1161\/jaha.113.000394"},{"key":"e_1_2_12_2_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgmr.2021.10080"},{"key":"e_1_2_12_2_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/ph16020320"},{"key":"e_1_2_12_2_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2018.06.00"},{"key":"e_1_2_12_3_2_1","unstructured":"Auray\u2010Blaiset\u00a0al. (2017). See above."}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.1087","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,11]],"date-time":"2024-12-11T13:22:49Z","timestamp":1733923369000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.1087"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6]]},"references-count":18,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2024,6]]}},"alternative-id":["10.1002\/cpz1.1087"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.1087","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6]]},"article-number":"e1087"}}