Quantification of Fatty Acid Profiles in Mice

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)].

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Han Y.M., Kim K.J., Jeong M., Park J.M., Go E.J., Kang J.X., Hong S.P, & Hahm K.B. (2016). Suppressed Helicobacter pylori-associated gastric tumorigenesis in Fat-1 transgenic mice producing endogenous ω-3 polyunsaturated fatty acids. Oncotarget, 7(41), 66606-66622.

Publication 2016

boron trifluoride (BF3)–methanol reagent hexane 6890 N Network GC System GC ChemStation Software

Boron trifluoride Colon Esters Fatty acid Flame ionization Gas chromatography Hexane Lipids Methanol Methylation Mice Nitrogen Powder Tails Tissue

Corresponding Organization : CHA Bundang Medical Center

Other organizations : Asan Medical Center, University of Ulsan, Harvard University, Massachusetts General Hospital

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Variable analysis

independent variables

Tissue type (mice tails or colon tissue)

dependent variables

Fatty acid profiles

control variables

Sample mass (1 cm of mice tails or 5 × 5 mm blocks of colon tissue)
Tissue grinding method (grounded to powder under liquid nitrogen)
Lipid extraction and fatty acid methylation protocol (heating at 100°C for 1 h under 14% boron trifluoride (BF3)–methanol reagent and hexane)
Gas chromatography settings (fully automated 6890 N Network GC System with flame-ionization detector)
Fatty acid identification (comparison with fatty acid standards)

controls

Positive control: Fatty acid standards (Nu-chek-Prep)
Negative control: Not explicitly mentioned

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