Pufa n 3

Total lipids from the lipid raft fractions from the hippocampus homogenates were extracted with chloroform/methanol (2:1 v/v) containing butylated hydroxytoluene (0.01%) as the antioxidant, according to Folch et al. (1957) [123 (link)] with small modifications. The organic solvent was evaporated under a stream of nitrogen, and the lipid content determined gravimetrically and stored in fresh solvent at −20 °C until further analysis. Lipids from the lipid rafts were subjected to acid-catalyzed transmethylation using 1 mL of toluene and 2 mL of 1% sulfuric acid (v/v) in methanol for 17 h in a heater at 50 °C. The resultant fatty acid methyl esters (FAMEs) and dimethylacetals (DMAs) were purified by thin layer chromatography (TLC), and quantified using a TRACE GC Ultra (Thermo Fisher Scientific, Waltham, MA, USA) gas chromatograph equipped with a flame ionization detector, with helium as the carrier gas. Individual FAME and DMA peaks were identified by referring to authentic standards (Mix C4-C24 and PUFA n° 3 from menhaden oil (Supelco Inc., Bellefonte, PA, USA).

Bovine serum albumin (BSA; fatty acid free), ALA, DHA, EPA, nonadecanoic acid (19:0), tridecanoic acid, gallic acid (GA), p-coumaric acid (PCA), syringic acid (SYA), butylated hydroxytoluene (BHT), potassium chloride and potassium bicarbonate were obtained from Sigma-Aldrich. Thin layer chromatography (TLC) (20×20 cm) plates, precoated with silica gel (without fluorescent indicator) were purchased from Macherey-Nagel (Düren, Germany). A fish oil standard (PUFA n-3) was supplied by Supelco (Supelco Park, Bellefonte, PA, USA). Delphinidin-3-O-glucoside (D3G), cyanidin-3-O-glucoside (C3G) and malvidin-3-O-glucoside (M3G) were obtained from Extrasynthese (Genay, France). Palm oil (PO) and rapeseed oil (RO) were obtained from William Hodgson & Co (Congleton, UK). All HPLC-grade solvents for chromatography and solvents for extraction (proanalysis quality) were purchased from Fisher Scientific (Fisher Scientific UK Ltd). The test polyphenolic compounds were dissolved in DMSO (Carl Roth, Karlsruhe, Germany), and stock solutions were stored at −80°C until usage.