Newer Approaches to the Isolation, Identification, and Quantitation of Steroids in Biological Materials

Sjövall, Jan; Axelson, Magnus

doi:10.1016/s0083-6729(08)61135-8

Cited by 61 publications

(19 citation statements)

References 532 publications

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“…Appropriate extraction and SPE procedures are required to set up reliable methods for the analysis of PREG-S and DHEA-S by GC-MS after hydrolysis. Alternatively, well-established methods based on lipophilic anion-exchange separations may be used to selectively isolate a steroid sulfate fraction (44). Therefore, our data and the study of Liu, Sjövall, and Griffiths (27) demonstrate that PREG-S and DHEA-S are not present at levels above 0.3 ng/g in rat brain.…”

Section: Discrepancies Between the Present And Previous Studiesmentioning

confidence: 57%

Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain

Lière

Pianos

Eychenne

et al. 2004

Journal of Lipid Research

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110

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A new sample preparation method coupled to GC-MS analysis was developed and validated for quantification of sulfate esters of pregnenolone (PREG-S) and dehydroepiandrosterone (DHEA-S) in rat brain. Using a solid-phase extraction recycling protocol, the results show that little or no PREG-S and DHEA-S ( Ͻ 1 pmol/g) is present in rat and mouse brain. These data are in agreement with studies in which steroid sulfates were analyzed without deconjugation. 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 (L-PREG and L-DHEA, respectively). These derivatives can be acylated very efficiently with heptafluorobutyric anhydride and triethylamine, and their levels in rodent brain ( ‫ف‬ 1 nmol/g) are much higher than those of their unconjugated counterparts. They are distinct from fatty acid esters, and preliminary data do not favor structures such as sulfolipids or sterol peroxides. Noncovalent interactions between steroids and proteolipidic elements, such as lipoproteins, could account for some experimental data. Given their abundance in rodent brain, the structural characterization and biological functions of L-PREG and L-DHEA in the central nervous system merit considerable attention. Four years ago, we developed and validated an analytical procedure for measuring trace amounts of neurosteroids in brain tissue by GC-MS (1). This method, which includes solid-phase extraction (SPE) and HPLC as fractionation and purification steps, is very suitable for quantifying simultaneously numerous neurosteroids in small individual regions of the central nervous system (CNS) (2) or peripheral nervous system (3) with high sensitivity and accuracy. Interest was primarily focused on pregnenolone (PREG), dehydroepiandrosterone (DHEA), and their sulfated conjugates (PREG-S and DHEA-S, respectively) and on progesterone (PROG) and allopregnanolone (3 ␣ -hydroxy-5 ␣ -pregnan-20-one).Neurosteroids are synthesized by glial cells and neurons from cholesterol or from blood-borne precursors (4). In particular, PREG-S and DHEA-S are known to be neuroactive, and their major effects are the modulation of several neuronal membrane receptors, such as ␥ -aminobutyric acid (5, 6), N -methyl-d -aspartate (7), and σ receptors, by affecting neuronal excitability and behavior (8, 9). Notably, a physiologic function of PREG-S was suggested by the positive correlation between PREG-S levels in the hippocampus of aged rats and their spatial memory performance (10). However, some studies do not favor the concept that PREG-S and DHEA-S are actually neurosteroids. Indeed, contradictory results have been obtained concerning the presence and activity of the hydroxysteroid sulfotransferase, implied in the sulfation of free steroid.

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Section: Discrepancies Between the Present And Previous Studiesmentioning

confidence: 57%

Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain

Lière

Pianos

Eychenne

et al. 2004

Journal of Lipid Research

Self Cite

110

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“…A temperature program of 500C-2700C with a rate of 380C/min was used. All analyses were performed in the selected ion monitoring mode for identification and quantitation of these steroids (13,14). Identities of the compounds listed in Table 1 …”

mentioning

confidence: 99%

Steroids and their conjugates in the mammalian brain.

Mathur

Prasad²,

Raju³

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

129

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Five steroids-38-hydroxypregn-S-en-20-one (pregnenolone; P), 3fi-hydroxy-5a-pregnan-20-one (3.3-AP), 3a-hydroxy-5a-pregnan-20-one (3a-AP), 3.-hydroxyandrost-5-en-17-one (dehydroepiandrosterone; D), and 3fi-hydroxy-5a-androstan-17-one (EpiA)-were extracted from the brains of adult male rats, rabbits, and dogs. The steroids exist in this organ as uncoajugated compounds and as sulfates, lipoidal esters, and sulfolipids. The techniques for separating these four classes ofsteroids from each other and for separating the five steroids from each other are described. In all cases, the steroids were identified by their retention time (Re) on HPLC, theirRt by gas chromatography, and by selected ion monitoring of their mass spectra. The latter were also used for quantification. In their reaction toward organic bases, the sulfolipid conjugates resemble previously described sulfolipids of cholesterol and sitosterol. These conijugates are relatively abundant in brain, particularly those of P and D, and this suggests that, in the search for the physiological significance of these brain constituents, these conjugates warrant attention.also seem to indicate that the above-mentioned steroids may be present in brain in amounts somewhat larger than those previously reported. Furthermore, until now only the sulfate and lipoidal esters of both P and D have been detected in the brain. Our findings reveal that another steroidal conjugate(s), still to be characterized, is also present. In particular, evidence is given here that indicates that steroidal conjugates comparable in nature to those that we previously described (11) for cholesterol and sitosterol are also present in substantial amounts in brain. We have designated this conjugated form a sulfolipid, with the possible structure StO-SO2-OL (where St is a steroid and L is a lipid moiety), because this nonpolar conjugate, when treated with an organic base such as triethylamine, can cleave to give either a product that behaves like the sulfuric acid ester of a steroid or an unconjugated steroid, like P. Although their exact structures have yet to be determined, the abundance of these conjugates in brain suggests that their relationship to brain function warrants corsideration.The most recent facet of steroid hormone biochemistry to evoke intense interest, astonishing activity, and discoveries that presage previously unforeseen scientific advances is that which deals with the relationship between steroids and neuronal-especially brain-function. One ofthe factors that has been responsible for stimulating the interest in this chapter of biochemistry has been the detection of steroids in brain (neurosteroids) by Baulieu and his colleagues (1). Another impetus for the development was the finding of Majewska et al. (2) that certain steroid derivatives are potent modulators of the Cl-channel associated with the type A receptor for y-aminobutyric acid. More recent findings have shown that other ion channels, particularly those associated with N-methyl-D-aspartate receptors, may also ...

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“…Analysis of sex steroids using gas-liquid chromatographic techniques and mass selective detection has been performed since the mid 1960s (see review of early studies by Sjovall & Axelson 1982). Routine analysis of sex steroids using mass spectrometry is a more recent occurrence with the advent of capillary chromatography columns and improvements in mass spectroscopy.…”

Section: Liquid/gas Chromatography With Mass Selective Detection (Lc/mentioning

confidence: 99%

Detailed Review Paper on Fish Screening Assays for the Detection of Endocrine Active Substances

2004

OECD Series on Testing and Assessment

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LIST OF TABLES ACKNOWLEDGMENTSThe OECD Secretariat would like to acknowledge the contributions of national experts who provided extensive comments on the draft version of this detailed review paper:• US Environmental Protection Agency, Office of Science Coordination and Policy, who prepared the original version of this document; • Brunel University, Department Biological Sciences, United Kingdom;• US Environmental Protection Agency, Mid-Continent Ecology Division (Duluth, MN);• University of Heidelberg, Zoological Institute, Aquatic Ecotoxicology, Germany;• Fraunhofer Institute for Molecular Biology and Applied Ecology (Schmallenberg) Germany;• ECT Ecotoxicology (Flörsheim), Germany;• Laboratory for Aquatic Organisms, Bayer CropScience (Monheim), Germany;• DHI Water & Environment, Environmental Risk Assessment (Hørsholm), Denmark, who assisted the OECD Secretariat in finalising the Detailed Review Paper; • Schering AG, Experimental Toxicology (Berlin), Germany;• University of Southern Denmark, Institute of Biology (Odenske), Denmark;• European Commission, ECB European Chemicals Bureau (Ispra) Italy;• Institut National de l'Environnement Industriel et des Risques (Verneuil-en-Halatte), France;• CEFAS -Centre for Environment Fisheries and Aquaculture Science, United Kingdom;• Department of the Army Corps of Engineers, United States;• Waterways Experiment Station, Environmental Laboratory (Vicksburg/MS, US). ENV/JM/MONO(2004)18 13 PREAMBLEIn 1998, a Task Force on Endocrine Disrupter Testing and Assessment (EDTA) was established at the request of OECD member countries. The EDTA Task Force is a Special Activity of the Test Guidelines Programme and its main objectives are to:identify the needs and prioritize the development of new and enhanced guidelines for the detection and characterization of endocrine disrupting chemicals;• develop a harmonized testing strategy for the screening and testing of endocrine disrupters;• manage validation work for newly developed and enhanced Test Guidelines as appropriate;and, • provide practical tools for sharing testing results and assessments.The need for new and updated test methods to detect and characterise endocrine disrupting chemicals has been expressed by the Task Force for the assessment of human health effects and environmental effects. At early meetings of the EDTA Task Force, it appeared that existing OECD Test Guidelines would insufficiently cover for endocrine-related effects, especially for the environment. Member countries decided to list test methods which could potentially cover effects of chemicals on the reproductive system (estrogen agonists/antagonists and androgen agonists/antagonists) and on the development (thyroid system), and proposed enhancements where needed. Fish screening assays have been developed for the detection of (anti-) estrogenic and (anti-)androgenic compounds in oviparous animals living in aquatic ecosystems, as there is good evidence that fish in the field are being impacted by endocrine active substances. This Detailed Review Paper (DRP) is...

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Newer Approaches to the Isolation, Identification, and Quantitation of Steroids in Biological Materials

Cited by 61 publications

References 532 publications

Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain

Novel lipoidal derivatives of pregnenolone and dehydroepiandrosterone and absence of their sulfated counterparts in rodent brain

Steroids and their conjugates in the mammalian brain.

Detailed Review Paper on Fish Screening Assays for the Detection of Endocrine Active Substances

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