title: Quantification of Dehydroepiandrosterone, 11-Deoxycortisol, 17-Hydroxyprogesterone, and Testosterone by Liquid Chromatography-Tandem Mass Spectrometry (LC/MS/MS).
abstract: Congenital adrenal hyperplasia (CAH) is a group of autosomal recessive disorders due to enzymatic defects in the biosynthetic pathway of cortisol and/or aldosterone. The analysis of cortisol, 17-hydroxyprogesterone (OHPG), dehydroepiandrosterone (DHEA), 11-deoxycortisol, and testosterone is generally performed in the diagnosis and/or follow-up of CAH. 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 17-hydroxyprogesterone, DHEA, 11-deoxycortisol, and testosterone. Stable-isotope labeled internal standards are added to serum samples and steroids are extracted by liquid-liquid extraction using 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.
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA.
authors.forename: Ada
authors.initials: A
authors.lastname: Munar
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA.
authors.forename: Clint
authors.initials: C
authors.lastname: Frazee
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, 2401 Gillham Road, Kansas City, MO, 64108, USA. ugarg@cmh.edu.
authors.forename: Uttam
authors.initials: U
authors.lastname: Garg
mesh_terms: D019326:17-alpha-Hydroxyprogesterone / Q000097:blood*; D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003350:Cortodoxone / Q000097:blood*; D003687:Dehydroepiandrosterone / Q000097:blood*; D013223:Statistics as Topic; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D013739:Testosterone / Q000097:blood*; D013997:Time Factors
chemical_list: D013739:Testosterone; D003687:Dehydroepiandrosterone; D019326:17-alpha-Hydroxyprogesterone; D003350:Cortodoxone
keywords: 11-Deoxycortisol; 17-Hydroxyprogesterone and testosterone; Dehydroepiandrosterone; Tandem mass spectrometry (LC/MS/MS)