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Updated: Oct 4, 2026
Semi-Targeted Ultra-High-Performance Chromatography Coupled to Mass Spectrometry Analysis of Phenolic Metabolites in Plasma of Elderly Adults
Published on: April 22, 2022
Determination of 17alpha-hydroxypregnenolone in human plasma by routine isotope dilution mass spectrometry using
S A Wudy1, M Hartmann, C Solleder
1Steroid Laboratory, Department of Pediatrics, University of Ulm Prittwitzstr. 43, D-89070, Ulm/Donau, Germany. stefan.wudy@paediat.med.uni-giessen.de
Abstract:
A first assay based on stable isotope dilution/gas chromatography-mass spectrometry has been developed for plasma 17alpha-hydroxypregnenolone, the leading marker of 3beta-hydroxysteroid dehydrogenase deficiency. Equilibration of plasma with internal standard was followed by solid phase extraction. After clean up using Sephadex LH-20 mini columns, heptafluorobutyrates were prepared as derivatives. Quantification was achieved by selected ion monitoring of m/z 467.0 (analyte) and m/z 471.0 (internal standard). 0.030 pmol of 17alpha-hydroxypregnenolone gave a signal to noise ratio of 3.7. Calibration plot was linear. Spiking experiments showed good accuracy with relative errors < 3.7%. Intraassay precision CV was 8.3% and interassay precision CV was 5.6%. Requiring small amounts of plasma, the rapid, convenient work up and the application of bench top GC/MS instrumentation proved our method suitable for routine clinical use in adults and children.
