Studies on Neurosteroids XXI: An Improved Liquid Chromatography–Tandem Mass Spectrometric Method for Determination of 5α-Androstane-3α,17β-diol in Rat Brains

Higashi, Tatsuya; Nishio, Takehiko; Yokoi, Hiroyuki; Ninomiya, Yasuaki; Shimada, Kazutake

doi:10.2116/analsci.23.1015

Cited by 16 publications

(4 citation statements)

References 17 publications

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“…To characterize the physiological function of 5a-androstane-3a,17b-diol (3a,5a-Adiol), a metabolite of testosterone (T), and the mechanism by which it affects the brain function, isonicotinoyl azide derivatization and LC-MS/MS were applied to analyze changes in the steroid levels by immobilization stress (Higashi et al, 2007a).…”

Section: Derivatization-based Lc-msmentioning

confidence: 99%

Enhancement of the capabilities of liquid chromatography–mass spectrometry with derivatization: General principles and applications

Zou

Liu

et al. 2011

Mass Spectrometry Reviews

135

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The integration of liquid chromatography-mass spectrometry (LC-MS) with derivatization is a relatively new and unique strategy that could add value and could enhance the capabilities of LC-MS-based technologies. The derivatization process could be carried out in various analytical steps, for example, sampling, storage, sample preparation, HPLC separation, and MS detection. This review presents an overview of derivatization-based LC-MS strategy over the past 10 years and covers both the general principles and applications in the fields of pharmaceutical and biomedical analysis, biomarker and metabolomic research, environmental analysis, and food-safety evaluation. The underlying mechanisms and theories for derivative reagent selection are summarized and highlighted to guide future studies.

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Section: Derivatization-based Lc-msmentioning

confidence: 99%

Enhancement of the capabilities of liquid chromatography–mass spectrometry with derivatization: General principles and applications

Zou

Liu

et al. 2011

Mass Spectrometry Reviews

135

View full text Add to dashboard Cite

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“…Although earlier studies on brain steroid concentrations analyzed by RIA or GC-MS have shown relatively high levels for these compounds, other reports based on more direct measurement using a LC-MS/MS method found substantially lower concentrations in brain tissue [9,39] [40][41][42][43][44]. The challenge for the LC-MS/MS quantification of unconjugated steroids rests on sensitivity and specificity.…”

Section: Introductionmentioning

confidence: 96%

Accurate and sensitive liquid chromatography/tandem mass spectrometry simultaneous assay of seven steroids in monkey brain

Bertin

Dury

et al. 2015

Steroids

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“…6,11) The mean steroid concentrations with standard deviations of the respective groups (nϭ10) are shown in Figs. 2a and b.…”

Section: Resultsmentioning

confidence: 99%

“…[6][7][8] Industries (Osaka, Japan). The derivatization reagents, isonicotinoyl azide (INA) 6) and 2-hydrazino-1-methylpyridine (HMP), 9) were synthesized in our laboratories. Strata-X (60 mg adsorbent) and Bond Elut Si (500 mg adsorbent) cartridges were purchased from Phenomenex (Torrance, CA, U.S.A.) and Varian (Harbor, CA, U.S.A.), respectively.…”

Section: Materials and Reagentsmentioning

confidence: 99%

Studies on Neurosteroids XXVI. Fluoxetine-Evoked Changes in Rat Brain and Serum Levels of Neuroactive Androgen, 5.ALPHA.-Androstane-3.ALPHA.,17.BETA.-diol

Higashi

Nagura

Shimada

et al. 2009

Biological & Pharmaceutical Bulletin

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Neuroactive steroids affect neurotransmission by action at the membrane ion-gated receptors and at other neurotransmitter receptors. 3a-Hydroxy-5a-pregnan-20-one (allopregnanolone, AP, Fig. 1), a representative neuroactive steroid, binds to the g-aminobutyric acid type A (GABA A ) receptors with a high affinity and positively modulates the action of GABA at these receptors, and hence elicits marked antidepressant, anxiolytic and anticonvulsant effects. 1) AP is synthesized from progesterone (PROG) via 5a-dihydroprogesterone (5a-DHP) by sequential reduction mediated by 5a-reductase and 3a-hydroxysteroid dehydrogenase (3a-HSD).Recent evidence also suggests that AP may be relevant to antidepressant drug action and depression pathophysiology.2,3) Fluoxetine (Fluox) is a typical selective serotonin (5-HT) reuptake inhibitor (SSRI) and is clinically used for the treatment of depression. However, Pinna et al. demonstrated that Fluox increases the brain AP content and subsequent activation of GABA A receptors at doses that are inactive toward the 5-HT reuptake, but does effectively treat depression. 3)Based on this result, Pinna et al. further stated that the term "SSRI" may be misleading in describing the pharmacological profile of Fluox and suggested the term "selective brain steroidogenic stimulant (SBSS)" as a better descriptor.3) Furthermore, rodent studies examining the Fluox-induced elevation in the brain AP level demonstrated that the effect is present to the same extent in both adrenalectomized and intact animals, suggesting that the Fluox-induced elevation in the brain AP level is due to de novo AP biosynthesis in the brain.2) 5a-Androstane-3a,17b-diol (3a,5a-Adiol, Fig. 1) is the best-characterized neuroactive androgen and derived from testosterone (T) by the action of 5a-reductase and 3a-HSD. 4) 3a,5a-Adiol is structurally similar to AP and is also a potent positive modulator of GABA A receptors with anxiolytic and anticonvulsant effects. 4,5) The anticonvulsant profile of 3a,5a-Adiol is highly consistent with AP, which have been proven by several animal experiments. 4) However, it is not currently known if Fluox influences the brain and serum levels of 3a,5a-Adiol. Examination of this point can be useful in evaluating the exact mechanisms of action of Fluox. Based on this background information, the objectives of this study are to determine the Fluox-evoked changes in the brain and serum 3a,5a-Adiol levels and to compare the level changes of 3a,5a-Adiol to those of AP. MATERIALS AND METHODS Materials and ReagentsAll the steroids including the deuterium-labeled internal standards (ISs) used in this study are those used in the previous studies.6-8) Each steroid was dissolved in and diluted with ethanol to prepare the standard solutions. Fluox was obtained from Wako Pure Chemical It is well known that fluoxetine (Fluox), a selective serotonin reuptake inhibitor, increases the brain content of allopregnanolone (AP), a potent neuroactive steroid that positively modulates the action of g g-aminobutyric acid (G...

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Studies on Neurosteroids XXI: An Improved Liquid Chromatography–Tandem Mass Spectrometric Method for Determination of 5α-Androstane-3α,17β-diol in Rat Brains

Cited by 16 publications

References 17 publications

Enhancement of the capabilities of liquid chromatography–mass spectrometry with derivatization: General principles and applications

Enhancement of the capabilities of liquid chromatography–mass spectrometry with derivatization: General principles and applications

Accurate and sensitive liquid chromatography/tandem mass spectrometry simultaneous assay of seven steroids in monkey brain

Studies on Neurosteroids XXVI. Fluoxetine-Evoked Changes in Rat Brain and Serum Levels of Neuroactive Androgen, 5.ALPHA.-Androstane-3.ALPHA.,17.BETA.-diol

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