To evaluate the saponifiable content of S. almeriensis, the biomass underwent a direct transesterification reaction, followed by gas chromatography (GC) analysis, as described by Jiménez Callejón et al. [68]. In brief, 10 mg of freeze-dried milled biomass were molturated with 10 mg of alumine for 5 min and stored at -21 °C until use. The molturated biomass was directly transesterified using 1 mL of acetyl chloride:methanol solution 1:20 vol/vol and 1 mL hexane. The reaction took place for 20 min at 105 °C and agitation. Afterwards, the mixture was left to reach room temperature, followed by the addition of 1 mL distil. water. The samples were then agitated and centrifuged. Two phases were formed, the upper one containing hexane and the produced FAMEs was removed and analyzed by GC. This was conducted in a Agilent Technologies 6890 N (Santa Clara, USA) GC, equipped with a capillary column of fused silica OmegaWax™ (0.25 mm x 30 m, 0.25 μm standard film, Supelco, Bellefonte, PA) and a flame ionization detector (FID). As carrier gas, nitrogen was used. Further technical details can be found in [68].

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