C11 0 fame

Seaweed fatty acids methyl esters (FAMEs) were prepared according to the methodology previously described by Kumari et al. [16] . Briefly, to 500 mg of powdered dried sample was added a mixture of 5 mL of acetyl chloride (Sigma-Aldrich®, Germany) and methanol (Fisher Scientific, UK) reagent in a ratio of 1:19 (v/v), spiked with 10 µL of an internal standard solution (1 mg/mL, C11:0 FAME (Sigma-Aldrich®, Germany)/C13:0 TAG (Sigma-Aldrich®, USA) in n-hexane (Merck KGaA, Germany)). The solution was esterified at 80 °C for 1 hour and then left at room temperature for cooling. Next, 1 mL of water and 2 mL of n-hexane were added to the mixture, vortexed, and centrifuged at 2057 × g for 5 min. The organic phase was collected, filtered, and dried using anhydrous sodium sulfate (Honeywell, Germany). Finally, solvents were later removed under a nitrogen flow and methyl esters were solubilized in 190 µL of n-hexane for GC-MS analysis.