All extraction and derivatization solvents were chromatography grade (EMD Chemicals, Billerica, MA). Monodeuterated ethanol was obtained from Acros Organics (New Jersey), the derivatizing reagent heptafluorobutyric acid anhydride (HFAA) was obtained from Fluka (Sigma-Aldrich, St. Louis, MO), and the neurosteroids pregnenolone, pregnanolone, 3α,5β-THDOC, and 3α,5β-androstanediol were obtained from Steraloids (Newport, RI). All other native neurosteroid standards were synthesized by the late Dr. Robert Purdy. Similar to Snelling et al. (2014) (link), the internal standard (ISTD) was 17β-estradiol-2,4,16,16,17-d5 (CDN Isotopes, Pointe-Claire, Quebec, Canada). Ultra-pure water (double distilled; ddH2O) from a MilliQ system (Millipore Corporation, Bedford, MA) was used. Activated powdered charcoal (Darco-G-60) and Dextran (T-70) were obtained from Sigma-Aldrich.
Protocol detail
Neurosteroid Quantification Protocol
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Monodeuterated ethanol
pregnenolone
pregnanolone
17β-estradiol-2,4,16,16,17-d5
MilliQ system
Darco-G-60
Dextran (T-70)
17β estradiol
Acid
Anhydride
Charcoal
Chromatography
Dextran
Ethanol
Heptafluorobutyric acid
Heptafluorobutyric anhydride
Isotopes
Neurosteroid
Pregnanolone
Pregnenolone
Solvents
Thdoc
Corresponding Organization :
Other organizations : Oregon Health & Science University
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Variable analysis
independent variables
- Extraction and derivatization solvents
- Levels of neurosteroids (pregnenolone, pregnanolone, 3α,5β-THDOC, and 3α,5β-androstanediol)
- Monodeuterated ethanol
- Derivatizing reagent heptafluorobutyric acid anhydride (HFAA)
- Neurosteroid standards
- Internal standard 17β-estradiol-2,4,16,16,17-d5
- Ultra-pure water (double distilled; ddH2O) from a MilliQ system
- Activated powdered charcoal (Darco-G-60)
- Dextran (T-70)
- Not explicitly mentioned
- Not explicitly mentioned
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