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 concentrations 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-D5, a stable isotope labeled analogue of DHEA, to real serum from healthy subjects. DHEA-D5 was 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.

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

CAMPI, BEATRICE
Co-primo
;
FRASCARELLI, SABINA
Co-primo
;
PAOLICCHI, ALDO;ZUCCHI, RICCARDO
Penultimo
;
SABA, ALESSANDRO
Ultimo
2018-01-01

Abstract

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 concentrations 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-D5, a stable isotope labeled analogue of DHEA, to real serum from healthy subjects. DHEA-D5 was 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.
2018
Campi, Beatrice; Frascarelli, Sabina; Pietri, Elisabetta; Massa, Ilaria; Donati, Caterina; Bozic, Roberto; Bertelloni, Silvano; Paolicchi, Aldo; Zucchi, Riccardo; Saba, Alessandro
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11568/875662
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