Eicosanoid Profiling in Colonic Mucosa

Eicosanoids were extracted from colonic scraped mucosa from TNBS-treated Fat-1 and Fads1 mice as previously described [47 (link), 48 (link), 53 ]. Liquid chromatography/tandem mass spectroscopic analyses were performed using a QuattroUltima mass spectrometer (Waters, Milford, MA, USA) equipped with an Agilent 1100 binary pump high-performance liquid chromatography system (Agilent Technologies, Santa Clara, CA, USA) according to a modified version of the method of Yang et al. [53 ]. Eicosanoids of interest were chromatographically separated using a Luna 3 μm phenyl-hexyl 4.6 × 100 mm analytic column (Phenomenex, Torrance, CA, USA) [48 (link)]. Eicosanoids were detected and quantified by multiple reaction mode monitoring of the transitions m/z as described elsewhere [48 (link), 54 (link)].