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Papers In Press, published online ahead of print September 1, 2004
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Unit 488, INSERM, Kremlin Bicetre 94276
Corresponding Author: liere{at}kb.inserm.fr
A new sample preparation method coupled to gas chromatography/mass spectrometry (GC/MS) analysis was developed and validated for quantification of sulfate esters of pregnenolone (PREG-S) and dehydroepiandrosterone (DHEA-S) in male rat brain. Using a solid-phase extraction (SPE) recycling protocol and a cleavage/acylation derivatization reaction, the results show that little or no PREG-S and DHEA-S (< 1 pmol/g) is present in rat and mouse brain. These data are in agreement with studies in which steroid sulfates are analyzed without deconjugation but previous experiments involving a hydrolysis step had indicated that substantial amounts of steroid sulfates were present in rodent brain. We suggest that the discrepancies between analyses with and without deconjugation are caused by internal contamination of brain extract fractions, supposed to contain steroid sulfates, by lipoidal forms of PREG and DHEA, designated L-PREG and L-DHEA, respectively. These derivatives can be acylated very efficiently with heptafluorobutyric anhydride (HFBA) and triethylamine (TEA) and their levels in rodent brain (roughly 1 nmol/g) are much higher than those of the unconjugated counterparts. They are distinct from fatty acid esters and preliminary data do not favor structures such as sulfolipids or sterol peroxides. Non-covalent interactions between steroids and proteolipidic elements, such as lipoproteins, could account for some experimental data. Given their abundance in rodent brain, structural characterization and biological functions of L-PREG and L-DHEA in the central nervous system (CNS) merit considerable attention.
Revised on August 31, 2004
Accepted on August 31, 2004
Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain
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