37 component standard

The fatty acid profile was determined through the use of a gas chromatograph coupled with an FID detector (Agilent Technologies 6850 Series II, Santa Clara, CA, USA) after trans-esterification with 2 N KOH in anhydrous methanol as described by Romano et al. [21 (link)]. The GC was equipped with a 90% biscyanopropyl−10% cyanopropyl phenyl siloxane capillary column (100 m, 0.25 mm, ID 0.20 µm) (Supelco, Bellefonte, PA, USA). The oven temperature schedule was 140 °C × 5 min, an increase of 4 °C/min to 175 °C for 20 min, and finally 3 °C/min 240 °C held for 10 min. The injector program temperature was 120 °C × 0.1 min, with an increase of 500 °C/min until reaching 260 °C held for 5 min. Helium was used as the carrier gas (flow rate 2 mL/min). The chromatogram peaks were identified using an external 37-component standard (Supelco, Bellefonte, PA, USA) by comparing the retention times of the standards with those of the samples under the same operating conditions. The results are expressed as relative % values.

Fatty acid methyl esters (FAMEs) concentrations were measured following transesterification using gas chromatography. The profile of fatty acids was assessed with a gas chromatograph equipped with an FID detector (Agilent Technologies 6850 Series II, Santa Clara, CA, USA) post-transesterification. The GC method of [53 ], with modification, was used. Briefly, 1 μL of the solution containing the FAMEs was injected into gas chromatograph equipped with a capillary column (100 m × 0.25 mm inner diameter, film thickness of 0.20 μm) with a polystationary phase (90% biscyanopropyl/10% cyanopropylphenyl siloxane) (Supelco, Bellafonte, PA, USA), hydrogen flame ionization detector (FID), and a programmed temperature vaporizer (PTV). Helium with a flow rate of 1 mL min⁻¹ was used as gas carrier. The oven temperature program was as follows: 140 °C × 5 min, a 4 °C min⁻¹ ramp to 175 °C for 20 min, and a 3 °C min⁻¹ ramp to 240 °C for 20 min. The chromatogram peaks were identified using an external 37-component standard (Supelco, Bellefonte, PA, USA). The results are expressed as relative % values.