A capillary gas chromatography/mass spectrometric method for the quantification of hydroxysteroids in human plasma

A specific and sensitive methodology for the quantitative determination of hydroxysteroids dehydroepiandrosterone and pregnenolone and their main metabolites in human plasma is described. Hydroxysteroids were extracted using methanol and steroids were further separated by reverse-phase high-performa...

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Bibliographic Details
Published in:Analytical biochemistry 2004, Vol.324 (1), p.123-130
Main Authors: Diallo, S, Lecanu, L, Greeson, J, Papadopoulos, V
Format: Article
Language:English
Subjects:
Analytical validation
Biotransformation
Chromatography, High Pressure Liquid
Dehydroepiandrosterone - blood
Dehydroepiandrosterone - metabolism
Gas Chromatography-Mass Spectrometry - methods
Gas chromatography/mass spectrometry
Humans
Hydroxysteroids - blood
Hydroxysteroids - chemistry
Molecular Structure
Neurosteroids
Plasma
Pregnenolone - blood
Pregnenolone - metabolism
Radioimmunoassay
Reproducibility of Results
Spectrophotometry
Steroids
Time Factors
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Summary:A specific and sensitive methodology for the quantitative determination of hydroxysteroids dehydroepiandrosterone and pregnenolone and their main metabolites in human plasma is described. Hydroxysteroids were extracted using methanol and steroids were further separated by reverse-phase high-performance liquid chromatography, allowing for minimization of the possible chromatographic interferences. Eluted fractions were collected, pooled, and analyzed by gas chromatography–mass spectrometry as trimethylsilyl ether derivatives. The quantification was performed with single-ion monitoring of the highly abundant m/ z 129 or m/ z 358 fragments. The combination of the chromatographic characteristics to the specific fragments ensured the selectivity and specificity of the method. Under these conditions the method was linear (typical R 2 is superior to 0.98 for all hydroxysteroids studied) over the concentration range of 2×10 −9 to 10 −6 M with good precision and accuracy.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2003.10.001