Quantification of dehydroepiandrosterone in human serum on a routine basis: development and validation of a tandem mass spectrometry method based on a surrogate analyte

In the clinical laboratories, dehydroepiandrostenedione (DHEA) is usually quantified by immunoassay-based methods, which are often affected by cross-reactivity with endogenous interferences, such as 4-androsten-3β-ol-17-one. The interfering compounds lead to a poor accuracy of the measurements, main...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2018, Vol.410 (2), p.407-416
Main Authors: Campi, Beatrice, Frascarelli, Sabina, Pietri, Elisabetta, Massa, Ilaria, Donati, Caterina, Bozic, Roberto, Bertelloni, Silvano, Paolicchi, Aldo, Zucchi, Riccardo, Saba, Alessandro
Format: Article
Language:English
Subjects:
Analytical Chemistry
Biochemistry
Calibration
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Chromatography, High Pressure Liquid - methods
Cross-reactivity
Dehydroepiandrosterone
Dehydroepiandrosterone - analogs & derivatives
Dehydroepiandrosterone - blood
Deuterium - analysis
Deuterium - blood
Food Science
Humans
Immunoassay
Ions
Kinetics
Laboratory Medicine
Limit of Detection
Liquid chromatography
Mass spectrometry
Mass spectroscopy
Monitoring/Environmental Analysis
Research Paper
Scientific imaging
Spectroscopy
Stable isotopes
Tandem Mass Spectrometry - methods
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Summary:In the clinical laboratories, dehydroepiandrostenedione (DHEA) is usually quantified by immunoassay-based methods, which are often affected by cross-reactivity with endogenous interferences, such as 4-androsten-3β-ol-17-one. The interfering compounds lead to a poor accuracy of the measurements, mainly at a low concentration level. The present paper describes a validated method based on tandem mass spectrometry coupled to liquid chromatography, for the accurate quantification of DHEA in serum. The peculiarity of this method is the use of calibrators and quality controls prepared by adding measured amounts of DHEA-D 5 , a stable isotope-labeled analogue of DHEA, to real serum from healthy subjects. DHEA-D 5 is used in place of DHEA, which is usually present in unstripped serum at physiological levels, as it has the same basic structure, provides an equivalent instrumental response, and can be easily distinguish by DHEA by mass spectrometry due to its different m / z value. The method proved to be sensitive, with a LLOD of 0.09 ng/mL and a LLOQ of 0.23 ng/mL, and selective, with overall performances that allow its use on a routine basis.
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-017-0731-x