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Routine method for the simultaneous quantification of 17 alpha-hydroxyprogesterone, testosterone, dehydroepiandrosterone, androstenedione, cortisol, and pregnenolone in human serum of neonates using gas chromatography-mass spectrometry

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dc.contributor.author Magnisali, P en
dc.contributor.author Dracopoulou, M en
dc.contributor.author Mataragas, M en
dc.contributor.author Dacou-Voutetakis, A en
dc.contributor.author Moutsatsou, P en
dc.date.accessioned 2014-06-06T06:48:15Z
dc.date.available 2014-06-06T06:48:15Z
dc.date.issued 2008 en
dc.identifier.issn 0021-9673 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/4037
dc.subject gas chromatography-mass spectrometry en
dc.subject 17 alpha-hydroxyprogesterone en
dc.subject testosterone en
dc.subject Delta 4-androstenedione en
dc.subject dehydroepiandrosterone en
dc.subject cortisol en
dc.subject pregnenolone en
dc.subject.classification Biochemical Research Methods en
dc.subject.classification Chemistry, Analytical en
dc.subject.other CONGENITAL ADRENAL-HYPERPLASIA en
dc.subject.other PROFILING STEROID-HORMONES en
dc.subject.other POLYCYSTIC-OVARY-SYNDROME en
dc.subject.other LIQUID-CHROMATOGRAPHY en
dc.subject.other HUMAN PLASMA en
dc.subject.other 21-HYDROXYLASE DEFICIENCY en
dc.subject.other CLINICAL-CHEMISTRY en
dc.subject.other QUALITY-CONTROL en
dc.subject.other WOMEN en
dc.subject.other IMMUNOASSAYS en
dc.title Routine method for the simultaneous quantification of 17 alpha-hydroxyprogesterone, testosterone, dehydroepiandrosterone, androstenedione, cortisol, and pregnenolone in human serum of neonates using gas chromatography-mass spectrometry en
heal.type journalArticle en
heal.language English en
heal.publicationDate 2008 en
heal.abstract Steroid determination by immunoassays results in significant interferences and inaccurate results. This study describes the development and validation of a new gas chromatographic-mass spectrometric method for the simultaneous quantification of 17 alpha-hydroxyprogesterone (17 alpha OHP), testosterone (T), dehydroepiandrosterone (DHEA), androstenedione (Delta 4-A), cortisol (F) and pregnenolone (Preg) in serum of neonates. Steroids were extracted and purified from 0.5 mL serum using diethyl ether and Extrelut mini NT1 column. The extracts were derivatized with N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA)/trimethylsilyl iodide (TMSI)/dithioerythritol (DTE) and the resulting trimethylsilyl derivatives were quantified by gas chromatography-selected ion monitoring-mass spectrometry (GC-SIM-MS). The detection limit for all steroids was lower than 0.1 ng/mL. The limit of quantification was 0.1 ng/mL for all steroids except cortisol which was at 0.25 ng/mL. d(3)-Testosterone and methyltestosterone served as internal standards. Precision for all compounds at the concentrations of 0.5, 1, 5 and 10 ng/mL (n = 10) in fortified steroid-free serum samples ranged from 0.8% to 16.6%. Accuracy was calculated at the concentrations of 0.5, 1, 5 and 10 ng/mL and ranged from -9.2% to 10.6% (n = 10). Linear calibration equations were obtained for all five steroids (0.125-31.25 ng/mL) and for cortisol (0.125-200 ng/mL). Relative recoveries at concentrations 1.0 and 12.5 ng/mL ranged from 70.5% to 97.5%. Absolute recoveries at the same concentrations ranged from 73.2% to 96.6%. Reference intervals were estimated for infants aged from 9 to 40 days. The proposed steroid profile is suitable for routine analysis and provides meaningful data for samples within normal range as well as those with elevated levels. (C) 2008 Elsevier B.V. All rights reserved. en
heal.publisher ELSEVIER SCIENCE BV en
heal.journalName JOURNAL OF CHROMATOGRAPHY A en
dc.identifier.issue 2 en
dc.identifier.volume 1206 en
dc.identifier.isi ISI:000259944000011 en
dc.identifier.spage 166 en
dc.identifier.epage 177 en


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