7αC4, 7α, 12α-dihydroxy-4-cholesten-3-one (7α12αC4) and 7α,12α-dihydroxy-5β- cholestan-3-one (7α12αC5β) were from Toronto Research Chemicals (Toronto, Ontario). 5α- Cholestanol was from Steraloids (Newport, RI) and epicoprostanol from Sigma-Aldrich (St. Louis, MO). BSTFA reagent was from Thermo Scientific (Bellefonte, PA). Human plasma and double charcoal stripped (DCS) plasma were from Golden West Bio (Temecula, CA). Methanol and water (GC-MS grade) were from Burdick and Jackson (Muskegon, MI). Formic acid (90%) was J.T.Baker brand, and glacial acetic acid (99.99%) was from Aldrich. Quaternary amonoxy (QAO) reagent (O-(3-trimethylammoniumpropyl) hydroxylamine) bromide is commercially available as Amplifex™ Keto reagent from http://www.sciex.com . The QAO-d3 reagent was provided by AB SCIEX.
Bstfa reagent
Manufactured by Thermo Fisher Scientific
BSTFA (N,O-Bis(trimethylsilyl)trifluoroacetamide) is a silylating agent used in gas chromatography-mass spectrometry (GC-MS) analysis. It is a common derivatization reagent that converts polar, non-volatile compounds into more volatile and thermally stable derivatives, enabling their analysis by GC-MS.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bstfa reagent
1
Cholesterol Metabolite Analysis Protocol
2
Sterol Quantification by GC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lipid extraction and GC-MS was conducted as previously described [59 (link)]. Pellets with 5x108 cells were used for lipid extraction. The dried total samples were resuspended in 100 μL chloroform added to 100 μL of BSTFA reagent (Thermo Scientific) and incubated at 70°C for 1 hour prior to GC-MS (Agilent 7890B GC–MS, Agilent 5977A MSD) analysis [60 (link)]. The retention time and mass spectral patterns of a sterol standard were used as references for lipid analysis. Sterol standards used in this study include cholesterol (Avanti 700100), ergosterol (Cayman 19850), 4,4-Dimethyl zymosterol (Avanti 700073), Zymosterol (Avanti 700068), and Eburicol (Smolecule S633611). The relative amount of sterols A and B were estimated based on the relative percentage of the sterol to ergosterol peak areas in each sample. Cholesterol was added as an internal standard for these analyses prior to lipid extraction.
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