Whatman no 3 filter paper

Indigenous Asian plants (Asparagus racemosus, Azadirachta indica, Cassia fistula, Curcuma aromatica, Datura metel, Piper retrofractum, Sargassum fusiforme, Senna tora, and Terminalia chebula), were selected based on their use in Asian traditional medicine as cognition enhancers. Extracts from all plants, with exception of Sargassum fusiforme, were prepared by the maceration method. All plants were dried in a hot air oven at 50 °C. The dried samples were finely powdered and soaked in 95% ethanol overnight at room temperature and filtered with Whatman filter paper No. 3 (Sigma-Aldrich, Bornem, Belgium). Three consecutive filtrates were pooled and evaporated in a vacuum rotary evaporator at 40 °C. The crude lipid fraction of Sargassum fusiforme was extracted using an adaptation of the Folch method^{65 (link)}. Briefly, Sargassum fusiforme was harvested by hand in early spring, washed in sea water, boiled for three hours and dried in a hot air oven. Next, dried Sargassum fusiforme (Clearspring Ltd, London, UK) was soaked overnight at room temperature in a 2:1 (v/v) chloroform and methanol (both VWR, Leuven, Belgium) mixture. The chloroform/methanol extract was evaporated under a N₂ stream and the remaining lipid fraction was dissolved in ethanol. Using gas chromatography/mass spectrometry, 24(S)-Saringosterol content of dried Sargassum fusiforme was determined to be 69.4 ng/mg.