{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T18:40:19Z","timestamp":1792089619005,"version":"4.4.0"},"reference-count":23,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100018952","name":"Birmingham Biomedical Research Centre","doi-asserted-by":"publisher","award":["NIHR203326"],"award-info":[{"award-number":["NIHR203326"]}],"id":[{"id":"10.13039\/501100018952","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Analytical Science Advances"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Steroids are critical for numerous physiological processes; disruption in their metabolism is associated with numerous endocrine disorders. Steroid quantification is essential to improve the understanding and diagnosis of these pathologies. Historically, urinary steroid profiling has been performed using low\u2010throughput gas chromatography mass spectrometry (GC\u2010MS), providing holistic coverage of steroid classes with low cross\u2010reactivity. Here, we translate our previous GC\u2010MS urinary steroid profile to a liquid chromatography tandem\u2010MS (LC\u2010MS\/MS) platform, offering a validated, comprehensive overview of steroid metabolism with comparatively low sample preparation times and increased throughput. Urinary steroids were enzymatically hydrolysed and extracted via C\n                    <jats:sub>18<\/jats:sub>\n                    solid\u2010phase extraction. Quantification was conducted using a triple quadrupole mass spectrometer. Chromatographic separation of 27 analytes was completed in 16 min using a C\n                    <jats:sub>18<\/jats:sub>\n                    \u2010T3 column. Due to chromatographic co\u2010elution of tetrahydrocortisol and 5\u03b1\u2010tetrahydrocortisol, a second injection was required on a BEH\u2010C\n                    <jats:sub>18<\/jats:sub>\n                    column for their separation. Lower limit of quantification (LLOQ) ranged from 2 to 20\u00a0ng\/mL, with accuracy (bias) ranging from \u201018.7% to 19.9%, and precision (percentage coefficient of variation [%CV]) ranging from 4.0% to 18.6%. Matrix effects remained within the ideal range &lt;\u00b115% for all steroids. Recovery ranged from 76% to 103%, and intra\u2010 and inter\u2010assay imprecision (CV) ranged from 0.8% to 14.9%. In 40 healthy volunteers, most analytes were detected above the LLOQ in over 95% of samples, although tetrahydro\u2010aldosterone (85%), 5\u2010pregenediol (68%), and pregnanetriol\u2010one (59%) demonstrated lower quantification rates. Diurnal and sex\u2010based variations were observed, with excretion levels significantly higher during daytime and in males. This robust, high\u2010throughput LC\u2010MS\/MS method facilitates the simultaneous quantification of multiple steroid classes, enhancing its utility for clinical and research applications in endocrine science.\n                  <\/jats:p>","DOI":"10.1002\/ansa.70087","type":"journal-article","created":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T01:00:57Z","timestamp":1777597257000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Comprehensive Quantitative Urinary Steroid Profiling of 29 Steroids Using Liquid Chromatography\u2010Tandem Mass Spectrometry"],"prefix":"10.1002","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-2983-9697","authenticated-orcid":false,"given":"Joshua T.","family":"Bain","sequence":"first","affiliation":[{"name":"Department of Metabolism and Systems Science, College of Medicine and Health University of Birmingham  Birmingham UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fozia","family":"Shaheen","sequence":"additional","affiliation":[{"name":"Department of Metabolism and Systems Science, College of Medicine and Health University of Birmingham  Birmingham UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Prete","sequence":"additional","affiliation":[{"name":"Department of Metabolism and Systems Science, College of Medicine and Health University of Birmingham  Birmingham UK"},{"name":"National Institute for Health and Care Research (NIHR) Birmingham Biomedical Research Centre  Birmingham UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorna C.","family":"Gilligan","sequence":"additional","affiliation":[{"name":"Department of Metabolism and Systems Science, College of Medicine and Health University of Birmingham  Birmingham UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5835-5643","authenticated-orcid":false,"given":"Angela E.","family":"Taylor","sequence":"additional","affiliation":[{"name":"Department of Metabolism and Systems Science, College of Medicine and Health University of Birmingham  Birmingham UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.JSBMB.2019.105439"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1210\/ER.2018\u201000262"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsbmb.2023.106445"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1210\/JC.2011\u20101565"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1210\/JC.2016\u20103285"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.CHROMA.2024.464735"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0378\u20104347(00)80683\u20100"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1039\/C7AN01990D"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1530\/EJE\u201015\u20100338"},{"key":"e_1_2_12_11_1","unstructured":"UK NEQAS for Steroid Hormones(Birmingham Quality 2025) https:\/\/birminghamquality.org.uk\/eqa\u2010programmes\/ster\/."},{"key":"e_1_2_12_12_1","unstructured":"Steroid Profiling by LC\u2010MS\/MS\u2014Chromsystems(Chromsystems 2025) https:\/\/chromsystems.com\/en\/products\/steroids.html."},{"key":"e_1_2_12_13_1","unstructured":"Steroid Panel LC\u2010MS | Immunoassays and MS(Tecan 2025) https:\/\/shop.tecan.com\/be\/en\/product\/30191875\/Steroid\u2010Panel\u2010LC\u2010MS."},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.CCA.2023.117301"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/JP952759G\/ASSET\/IMAGES\/LARGE\/JP952759GF00009.JPEG"},{"key":"e_1_2_12_16_1","unstructured":"ICH M10 on bioanalytical method validation\u2014Scientific guideline | European Medicines Agency (EMA) https:\/\/www.ema.europa.eu\/en\/ich\u2010m10\u2010bioanalytical\u2010method\u2010validation\u2010scientific\u2010guideline."},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.JPBA.2018.11.035"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.D119000588"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/RCM.9719;CTYPE:STRING:JOURNAL"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/BMC.4723"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1373\/CLINCHEM.2017.271106"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.JPBA.2023.115785"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.1210\/EN.2016\u20101297"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1515\/cclm\u20102023\u20100250"}],"container-title":["Analytical Science 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