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The study aimed to establish assay-specific cut-off limits for cortisol after adrenocorticotropic hormone (ACTH) stimulation, comparing Roche Elecsys Cortisol II immunoassay to liquid chromatography-mass spectrometry (LC-MS\/MS), and to assess the impact of patient characteristics, estrogen containing oral contraceptives as well as relation to other adrenocortical steroid hormone dynamics.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>One hundred healthy participants underwent a 250\u2009\u03bcg ACTH-test, with plasma samples analyzed using ElecsysCortI, ElecsysCortII, and LC-MS\/MS. Cortisone, corticosterone, 17-OH-progesterone, dehydroepiandrosterone sulfate (DHEAS), androstenedione, and testosterone were additionally analyzed with LC-MS\/MS. Cut-off limit for a normal cortisol response to the ACTH-test was defined as: 2.5th percentile\u20131.96\u2009\u00d7\u2009SE.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>ElecsysCort II measured cortisol concentrations 21% (95% CI: 19\u201322%) lower than ElecsysCort I. Cut-off limits for cortisol 30 and 60\u2009min after ACTH were 426 and 485\u2009nmol\/L (ElecsysCort II) and 411 and 470\u2009nmol\/L (LC-MS\/MS). Cut-offs were unaffected by gender, or body-composition. The ACTH-test resulted in significantly increased adrenocortical steroid hormones, except for decreased cortisone concentrations (both sexes), and decreased testosterone in men (1.9\u2009nmol\/L, 95% CI: 1.3\u20132.5). Testosterone was increased in women (0.07\u2009nmol\/L, 95% CI: 0.02\u20130.13).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>ElecsysCort II has high analytical performance and yields significantly lower cortisol concentrations than prior polyclonal immunoassays. This clinically relevant difference underscores the necessity for revised cut-off limits for improved diagnostic precision. Suggested 30-minute cortisol cutoff limits are 411\u2009nmol\/L (LC-MS\/MS) and 426\u2009nmol\/L (ElecsysCort II). Adrenocortical steroids increased upon ACTH stimulation, except for cortisone in both sexes and testosterone in men, both of which decreased.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s12020-024-03752-0","type":"journal-article","created":{"date-parts":[[2024,3,9]],"date-time":"2024-03-09T03:01:16Z","timestamp":1709953276000},"page":"321-330","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Determination of cortisol cut-off limits and steroid dynamics in the ACTH stimulation test: a comparative analysis using Roche Elecsys Cortisol II immunoassay and LC-MS\/MS"],"prefix":"10.1007","volume":"85","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-8808-2123","authenticated-orcid":false,"given":"Sema","family":"Okutan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nanna Thurmann","family":"J\u00f8rgensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lars Engers","family":"Pedersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3939-3388","authenticated-orcid":false,"given":"Stina Willemoes","family":"Borresen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda","family":"Hilsted","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lennart Friis","family":"Hansen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5903-3355","authenticated-orcid":false,"given":"Ulla","family":"Feldt-Rasmussen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6075-0821","authenticated-orcid":false,"given":"Marianne","family":"Klose","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,3,9]]},"reference":[{"key":"3752_CR1","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1111\/cen.12528","volume":"82","author":"Y Simsek","year":"2015","unstructured":"Y. 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