{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T10:36:33Z","timestamp":1781951793665,"version":"3.54.5"},"reference-count":19,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T00:00:00Z","timestamp":1747872000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Background: Urinary steroid profiling plays a key role in the diagnosis of inherited and acquired endocrine disorders. Despite the proven diagnostic value of gas chromatography\u2013mass spectrometry (GC-MS), standardized and clinically validated protocols for extended steroid panels remain limited. Methods: We developed and validated a GC-MS method for the quantification of 32 urinary steroid metabolites, including androgens, estrogens, progestins, glucocorticoids, and mineralocorticoids. Sample preparation involved solid-phase extraction, enzymatic hydrolysis, and dual derivatization, followed by chromatographic separation and mass detection under full scan mode. Validation followed ICH M10 guidelines. Results: The method demonstrated high selectivity, accuracy (within \u00b115%), and precision (CV% &lt; 15%) across three QC levels. Limits of Quantification were estimated using the Hubaux\u2013Vos approach and were suitable for detecting both physiological and pathological steroid concentrations. Robustness and matrix effect tests confirmed the method\u2019s reliability and reproducibility. Conclusions: This GC-MS protocol enables comprehensive urinary steroid profiling and calculation of diagnostic ratios for inborn errors of steroid metabolism and endocrine disorders. The method is suitable for clinical application and future integration into personalized medicine workflows.<\/jats:p>","DOI":"10.3390\/life15060829","type":"journal-article","created":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T04:32:37Z","timestamp":1747888357000},"page":"829","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Steroidomics via Gas Chromatography\u2013Mass Spectrometry (GC-MS): A Comprehensive Analytical Approach for the Detection of Inborn Errors of Metabolism"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2585-9607","authenticated-orcid":false,"given":"Francesco","family":"Chiara","sequence":"first","affiliation":[{"name":"Department of Physics, University of Trento, 38123 Povo, Italy"},{"name":"Department of Clinical and Biological Sciences, University of Turin, 10043 Orbassano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4933-0011","authenticated-orcid":false,"given":"Sarah","family":"Allegra","sequence":"additional","affiliation":[{"name":"Department of Clinical and Biological Sciences, University of Turin, 10043 Orbassano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8843-2315","authenticated-orcid":false,"given":"Simona","family":"Liuzzi","sequence":"additional","affiliation":[{"name":"Clinical Biochemistry Laboratory \u201cBaldi e Riberi\u201d, A.O.U. Citt\u00e0 della Salute e della Scienza di Torino, 10134 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6719-1946","authenticated-orcid":false,"given":"Maria Paola","family":"Puccinelli","sequence":"additional","affiliation":[{"name":"Clinical Biochemistry Laboratory \u201cBaldi e Riberi\u201d, A.O.U. Citt\u00e0 della Salute e della Scienza di Torino, 10134 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0431-8369","authenticated-orcid":false,"given":"Giulio","family":"Mengozzi","sequence":"additional","affiliation":[{"name":"Clinical Biochemistry Laboratory \u201cBaldi e Riberi\u201d, A.O.U. Citt\u00e0 della Salute e della Scienza di Torino, 10134 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9841-7802","authenticated-orcid":false,"given":"Silvia","family":"De Francia","sequence":"additional","affiliation":[{"name":"Department of Clinical and Biological Sciences, University of Turin, 10043 Orbassano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1093\/clinchem\/17.8.802","article-title":"Metabolic profiles: Gas-phase methods for analysis of metabolites","volume":"17","author":"Horning","year":"1971","journal-title":"Clin. 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