Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) Method for the Quantification of Steroids Androstenedione, Dehydroepiandrosterone, 11-Deoxycortisol, 17-Hydroxyprogesterone, and Testosterone.

Document Type

Article

Publication Date

2022

Identifier

DOI: 10.1007/978-1-0716-2565-1_40

Abstract

Congenital adrenal hyperplasia (CAH) is a group of autosomal-recessive disorders due to deficiency of 11- or 21-hydroxylase. The analysis of cortisol, androstenedione, 17-hydroxyprogesterone (OHPG), dehydroepiandrosterone (DHEA), 11-deoxycortisol, and testosterone is generally performed in the diagnosis and/or follow-up of CAH. Analysis of specific steroids is also performed in other disorders such as evaluation of hirsutism or infertility in females and hypogonadism in males. Cortisol is generally analyzed by immunoassays, whereas other hormones are preferably assayed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A multiple reaction monitoring, positive mode atmospheric pressure chemical ionization, LC-MS/MS method is described for the simultaneous quantification of androstenedione, 17-hydroxyprogesterone, DHEA, 11-deoxycortisol and testosterone. The method involves addition of labeled internal standards to serum samples and extraction of steroids in methyl tert-butyl ether. The extract is evaporated under stream of nitrogen, and the residue is reconstituted in methanol and analyzed by LC-MS/MS.

Journal Title

Methods in molecular biology (Clifton, N.J.)

Volume

2546

First Page

451

Last Page

457

MeSH Keywords

17-alpha-Hydroxyprogesterone; Androgens; Androstenedione; Chromatography, Liquid; Cortodoxone; Dehydroepiandrosterone; Female; Humans; Hydrocortisone; Male; Methanol; Steroid 21-Hydroxylase; Steroids; Tandem Mass Spectrometry; Testosterone

Keywords

11-Deoxycortisol; 17-Hydroxyprogesterone and testosterone; Androstenedione; Dehydroepiandrosterone; Tandem mass spectrometry (LC-MS/MS)

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