4.6. GC-FID Analysis of FAME

Acid catalyzed transesterification with 0.5 N H₂SO₄ in methanol was used to produce FAME of recovered lipid classes [44]. FAME were determined using a GC (Bruker, SCION 436-GC) fitted with a DB-23 column ((50%-cyanopropyl)-methylpolysiloxane) (30 m × 0.25 m × 0.25 µm film thickness, Agilent Technologies, Santa Clara, CA, USA) and a flame ionization detector (FID). Splitless injection was used with an injector temperature of 250 °C. FAME samples were separated using the following temperature program: the initial temperature was held at 60 °C for 1 min, then increased to 153 °C at 45 °C min⁻¹, held for 2 min, then increased to 174 °C at 2.3 °C min⁻¹ and held for 0.2 min, increased to 205 °C at 2.5 °C min⁻¹ and held for 2.5 min for a total run time of 41 min.