{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T10:59:01Z","timestamp":1781607541120,"version":"3.54.5"},"reference-count":39,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,6]],"date-time":"2024-05-06T00:00:00Z","timestamp":1714953600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI19\/00774"],"award-info":[{"award-number":["PI19\/00774"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PID2022-138243OB-I00"],"award-info":[{"award-number":["PID2022-138243OB-I00"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"],"award-info":[{"award-number":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["2021 SGR 00881"],"award-info":[{"award-number":["2021 SGR 00881"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Agencia Estatal de Investigaci\u00f3n","award":["PI19\/00774"],"award-info":[{"award-number":["PI19\/00774"]}]},{"name":"Agencia Estatal de Investigaci\u00f3n","award":["PID2022-138243OB-I00"],"award-info":[{"award-number":["PID2022-138243OB-I00"]}]},{"name":"Agencia Estatal de Investigaci\u00f3n","award":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"],"award-info":[{"award-number":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"]}]},{"name":"Agencia Estatal de Investigaci\u00f3n","award":["2021 SGR 00881"],"award-info":[{"award-number":["2021 SGR 00881"]}]},{"DOI":"10.13039\/501100002809","name":"FEDER","doi-asserted-by":"publisher","award":["PI19\/00774"],"award-info":[{"award-number":["PI19\/00774"]}],"id":[{"id":"10.13039\/501100002809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002809","name":"FEDER","doi-asserted-by":"publisher","award":["PID2022-138243OB-I00"],"award-info":[{"award-number":["PID2022-138243OB-I00"]}],"id":[{"id":"10.13039\/501100002809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002809","name":"FEDER","doi-asserted-by":"publisher","award":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"],"award-info":[{"award-number":["MCIN\/AEI\/10.13039\/501100011033\/FEDER UE"]}],"id":[{"id":"10.13039\/501100002809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002809","name":"FEDER","doi-asserted-by":"publisher","award":["2021 SGR 00881"],"award-info":[{"award-number":["2021 SGR 00881"]}],"id":[{"id":"10.13039\/501100002809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Introduction: Urine free cortisol measurements are routinely performed to evaluate hypercortisolism. Despite their analytical inaccuracy, immunoassay-based methods are frequently used. Advances in liquid chromatography\u2013high-resolution mass spectrometry (LC-HRMS) facilitate the incorporation of powerful diagnostic tools into clinical laboratories. In addition to its high analytical specificity and simultaneous analysis of different metabolites, accurate mass measurement allows for untargeted compound identification, which may help to identify clinically relevant metabolites or drugs. Methods: The present study aimed to validate a simple routine LC\u2013HRMS method to quantify cortisol, cortisone, 6\u03b2-hydroxycortisol, and 18-hydroxycortisol simultaneously in human urine. Additionally, the study also validated a GC-MS method for the same steroids, evaluated their cross-reactivity with commercial cortisol immunoassays, and quantified the 24 h urine excretion in patients under clinical suspicion or follow-up for hypercortisolism. Results: The LC-HRMS method involved liquid\u2013liquid extraction using dichloromethane, micro-LC for chromatographic separation and detection using the accurate masses of the steroids, and simultaneous high-resolution full scan acquisition. The method presented acceptable linearity, precision, and accuracy. Significant interference from 6\u03b2-hydroxycortisol and cortisone was demonstrated in the cortisol immunoassays, which impacted their reliability in the follow-up of patients with hypercortisolism and significant changes in these cortisol metabolites (i.e., due to drug-induced changes in CYP3A4 activity). Conclusion: A rapid and accurate routine LC-HRMS method was validated, which is useful for the evaluation of hypercortisolism and other disorders of glucocorticoid and mineralocorticoid metabolism.<\/jats:p>","DOI":"10.3390\/biom14050558","type":"journal-article","created":{"date-parts":[[2024,5,6]],"date-time":"2024-05-06T15:05:01Z","timestamp":1715007901000},"page":"558","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["LC-HRMS and GC-MS Profiling of Urine Free Cortisol, Cortisone, 6\u0392-, and 18-Hydroxycortisol for the Evaluation of Glucocorticoid and Mineralocorticoid Disorders"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3271-1371","authenticated-orcid":false,"given":"Gregori","family":"Casals","sequence":"first","affiliation":[{"name":"Biochemistry and Molecular Genetics Department, Hospital Cl\u00ednic of Barcelona, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 08036 Barcelona, Spain"},{"name":"Department of Fundamental and Clinical Nursing, Faculty of Nursing, University of Barcelona, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Antonieta","family":"Ballesteros","sequence":"additional","affiliation":[{"name":"Service of Clinical Analysis, Hospital Universitario Son Espases, 07210 Palma de Mallorca, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angielys","family":"Zamora","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, Hospital General Universitario Gregorio Mara\u00f1\u00f3n, 28007 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Genetics Department, Hospital Cl\u00ednic of Barcelona, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7776-3300","authenticated-orcid":false,"given":"Guillermo","family":"Fern\u00e1ndez-Varo","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Genetics Department, Hospital Cl\u00ednic of Barcelona, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mireia","family":"Mora","sequence":"additional","affiliation":[{"name":"Department of Endocrinology and Nutrition, Hospital Clinic, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain"},{"name":"Department of Medicine, Faculty of Medicine and Health Science, University of Barcelona, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felicia A.","family":"Hanzu","sequence":"additional","affiliation":[{"name":"Department of Endocrinology and Nutrition, Hospital Clinic, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain"},{"name":"Department of Medicine, Faculty of Medicine and Health Science, University of Barcelona, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9074-2272","authenticated-orcid":false,"given":"Manuel","family":"Morales-Ruiz","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Genetics Department, Hospital Cl\u00ednic of Barcelona, Institut d\u2019Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Centro de Investigaci\u00f3n Biom\u00e9dica en Red de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 08036 Barcelona, Spain"},{"name":"Department of Biomedicine, Faculty of Medicine and Health Science, University of Barcelona, 08036 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1093\/clinchem\/46.6.793","article-title":"How much \u201cUFC\u201d is really cortisol?","volume":"46","author":"Murphy","year":"2000","journal-title":"Clin Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1016\/j.beem.2013.10.008","article-title":"Determination of cortisol in serum, saliva and urine","volume":"27","author":"Turpeinen","year":"2013","journal-title":"Best. 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