Boron trifluoride bf3 methanol reagent

Fatty acid profiles were analyzed using gas chromatography as described previously [19 (link)]. Briefly, 1 cm of mice tails (in order to perform the phenotyping of mice) or blocks of colon tissue (5 × 5 mm) were grounded to powder under liquid nitrogen. Samples were then subjected to extraction of total lipids and fatty acid methylation by heating at 100°C for 1 h under 14% boron trifluoride (BF3)–methanol reagent (Sigma, St. Louis, MO) and hexane (Sigma). Fatty acid methyl esters were analysed by gas chromatography using a fully automated 6890 N Network GC System (Agilent Technologies) equipped with a flame-ionization detector. Peaks of resolved fatty acids were identified by comparison with fatty acid standards (Nu-chek-Prep), and area percentage for all resolved peaks was analysed using GC ChemStation Software (Agilent Technologies, Santa Clara, CA) [61 (link)].

The profile of FAs in the heart was determined by gas chromatography as described previously.16, 17 In brief, ~10 mg heart tissue was ground into powder under liquid nitrogen, then centrifuged for 5 minutes at 100 g. The sediment was retained for FA methylation by 14% boron trifluoride (BF3)–methanol reagent (Sigma‐Aldrich, St Louis, MO, USA) at 100°C for 1 hour. FA methyl esters were analysed with the Agilent HP6890N gas chromatography system equipped with a flame ionization detector (Agilent, Palo Alto, CA, USA).FA peaks were identified by comparing their relative retention times with commercial mixed standards (Nu‐Chek Prep, Elysian, MN, USA), and the area percentages for all resolved peaks were analysed using GC Chemstation software. FA mass was determined by comparing areas of various analysed FAs to that of a fixed concentration of external standard.

Fatty acid profiles of mouse diets and tail and colon tissues were analyzed by gas chromatography (GC), as described previously [18 (link),97 (link)]. Briefly, tissue or food samples were ground to powder under liquid nitrogen and subjected to total lipid extraction and fatty acid methylation by 14% boron trifluoride (BF3)-methanol reagent (Sigma-Aldrich) at 100 °C for 1 h. Fatty acid methyl esters were analyzed using a fully automated HP5890 gas chromatography system equipped with a flame-ionization detector (Agilent Technologies, Palo Alto, CA, USA). The fatty acid peaks were identified by comparing their relative retention times with the mixed commercial standards (NuChek Prep, Elysian, MN, USA), and the area percentage for all resolved peaks were analyzed by using a PerkinElmer M1 integrator. The fatty acids examined as total n-6 PUFA with GC include: Linoleic acid (C18:2n6), Gamma-linolenic acid (C18:3n6), Eicosadienoic acid (C20:2n6), Dihomo-gamma-linolenic acid (C20:3n6), Arachidonic acid (C20:4n6), Docosadienoic acid (C22:2n6), Adrenic acid (C22:4n6) and Docosapentaenoic acid (22:5n6). The fatty acids examined as total n-3 PUFA with GC include: α-Linolenic acid (C18:3n3), Eicosatrienoic acid (C20:3n3), Eicosapentaenoic acid (C20:5n3), Docosapentaenoic acid (C22:5n3), and Docosahexaenoic acid (C22:6n3).