Simultaneous measurement of 17 endogenous steroid hormones in human serum by liquid chromatography-tandem mass spectrometry without derivatization.
J Steroid Biochem Mol Biol
; 243: 106578, 2024 Jul 04.
Article
em En
| MEDLINE
| ID: mdl-38971335
ABSTRACT
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⯵L serum. The targeted steroids for quantification included androgens dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), dihydrotestosterone (DHT), 11-oxyandrogens 11ß-hydroxy-androstenedione (11OHA4), 11-keto-androstenedione (11KA4), 11ß-hydroxy-testosterone (11OHT), 11-keto-testosterone (11KT), progestogens 17α-hydroxy-progesterone (17OHP4), progesterone (P4), 11ß-hydroxy-progesterone (11OHP4), 11-keto-progesterone (11KP4), mineralocorticoids aldosterone, corticosterone, and glucocorticoids 11-deoxycortisol, cortisol, and cortisone. The lower limits of quantification (LLOQ) were 0.05â¯ng/mL for A4, T, 11KA4, P4, and cortisone, 0.1â¯ng/mL for DHT, 11OHA4, 11OHT, 11KT, 17OHP4, 11OHP4, 11KP4, corticosterone, aldosterone, 11-deoxycortisol, and cortisol, and 0.5â¯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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Idioma:
En
Revista:
J Steroid Biochem Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
/
BIOQUIMICA
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Eslovênia