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Medientyp:
E-Artikel
Titel:
Determination of 17-Hydroxyprogesterone in Plasma by Stable Isotope Dilution/Benchtop Liquid Chromatography-Tandem Mass Spectrometry
Beteiligte:
Wudy, Stefan A.;
Hartmann, Michaela;
Svoboda, Michal
Erschienen:
S. Karger AG, 2000
Erschienen in:
Hormone Research in Paediatrics, 53 (2000) 2, Seite 68-71
Sprache:
Englisch
DOI:
10.1159/000023516
ISSN:
1663-2818;
1663-2826
Entstehung:
Anmerkungen:
Beschreibung:
An assay based on stable isotope dilution liquid chromatography-tandem mass spectrometry (ID/LC-MS-MS) was developed for the quantification of 17-hydroxyprogesterone, the most important indicator of 21-hydroxylase deficiency in human plasma. Plasma was extracted using ethyl acetate and Extrelut<sup>®</sup> columns. LC was performed on a reversed-phase C18 column using a water/methanol gradient. A benchtop triple quadrupole mass spectrometer, operating in selected reaction monitoring mode, served as mass detector. The analytical run time was 9 min per sample. The sensitivity was high: 0.06 pmol of 17-hydroxyprogesterone yielded a signal-to-noise ratio of 13. Precision (CV) and accuracy (relative error) derived from the analyses of unspiked and spiked validation samples were 7.4–12.0% and 6.4%, respectively. When analyzing the same samples – median (range), in nanomoles per liter – from neonates and adults independently by ID/LC-MS-MS as well as by ID/gas chromatography (GC)-MS, corresponding results were obtained: neonates (n = 10), ID/LC-MS-MS 3.99 (0.48–16.05), ID/GC-MS 5.39 (1.57–13.02); adults (n = 10), ID/LC-MS-MS 2.66 (1.39–6.15), ID/GC-MS 2.54 (0.51–5.12). The technique permitted reliable detection of classical and nonclassical forms of 21- hydroxylase deficiency. The much simpler sample preparation, the faster analytical run time and the operational ease possible with ID/LC-MS-MS permit a considerable increase of sample testing per day without compromising on analytical sensitivity and specificity. We expect that benchtop tandem mass spectrometry will open new avenues in clinical steroid analysis.