Simultaneous measurement of 17 endogenous steroid hormones in human serum by liquid chromatography-tandem mass spectrometry without derivatization

Gjorgoska, Marija (author), Lanišnik-Rižner, Tea (author)

Mass spectrometric-based steroidomics is a valuable analytical approach that gives a comprehensive understanding of the interlinked steroid biosynthetic pathways. Here, we describe a rapid and versatile liquid chromatography-tandem mass spectrometry (LC-MS/MS) method designed to accurately quantify endogenous steroids in human serum. Sample preparation involved liquid-liquid extraction with methyl tert-butyl ether (MTBE) from 180 µL serum. The targeted steroids for quantification included androgens: dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), dihydrotestosterone (DHT), 11-oxyandrogens: 11β-hydroxy-androstenedione (11OHA4), 11-keto-androstenedione (11KA4), 11β-hydroxy-testosterone (11OHT), 11-keto-testosterone (11KT), progestogens: 17α-hydroxy-progesterone (17OHP4), progesterone (P4), 11β-hydroxy-progesterone (11OHP4), 11-keto-progesterone (11KP4), mineralocorticoids: aldosterone, corticosterone, and glucocorticoids: 11-deoxycortisol, cortisol, and cortisone. The lower limits of quantification (LLOQ) were 0.05 ng/mL for A4, T, 11KA4, P4, and cortisone, 0.1 ng/mL for DHT, 11OHA4, 11OHT, 11KT, 17OHP4, 11OHP4, 11KP4, corticosterone, aldosterone, 11-deoxycortisol, and cortisol, and 0.5 ng/mL for DHEA. Accuracy, precision, reproducibility, and recovery fell within acceptable limits for bioanalytical method validation. Using serum samples from 29 premenopausal women in different menstrual phases, we demonstrated the clinical utility of our method, which showed sufficient sensitivity to reliably quantify all targeted steroids at levels typically found in circulation, except for 11OHP4 and 11KP4.

• Language: English
• Keywords: liquid chromatography-tandem mass spectrometry, androgens, progestogens, mineralocorticoids, glucocorticoids, 11-oxygenenated steroids
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: 8 str.
• Numbering: Vol. 243, art. 106578
• PID: 20.500.12556/RUL-159798
• UDC: 577.2
• ISSN on article: 1879-1220
• DOI: 10.1016/j.jsbmb.2024.106578
• COBISS.SI-ID: 202258435
• Publication date in RUL: 25.07.2024

Record is a part of a journal

• Title: The journal of steroid biochemistry and molecular biology
• Shortened title: J. steroid biochem. mol. biol.
• Publisher: Elsevier
• ISSN: 1879-1220
• COBISS.SI-ID: 57401603

Licences

• License: CC BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International
• Link: http://creativecommons.org/licenses/by-nc/4.0/
• Description: A creative commons license that bans commercial use, but the users don’t have to license their derivative works on the same terms.

Secondary language

• Language: Slovenian
• Keywords: tekočinska kromatografija s tandemsko masno spektrometrijo, androgeni, gestageni, mineralokortikoidi, glukokortikoidi, 11-oksigenirani steroidi

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: J3-2535
• Name: Vloga androgenov pri hormonsko odvisnih boleznih: pomen za diagnostiko in zdravljenje

• Funder: ARRS - Slovenian Research Agency
• Project number: P3-0449
• Name: Translacijska molekularna endokrinologija za zdravje žensk