2.6 Extraction of fatty alcohols from small-scale samples
In small-scale screening experiments, 1 mL of cell culture was sampled into 4 mL glass vials and centrifuged at 3,500 g, 5 min, 21°C. The supernatant was removed, and 1 mL of ethyl acetate:ethanol mixture (85:15, v/v) was added to the pellet together with 10 µL of internal standard (2 g/L of methyl nonadecanoate (19:Me) in ethyl acetate). Vials were vortexed for 20 s and incubated for 1 h, followed by 5 min vortexing. 300 µL of water was added, samples vortexed for 10 s, centrifuged at 3,500 g, 5 min, 21°C, and the upper organic layer was taken for analysis.
Samples from bioreactors were processed as follows. 100 µL of cell culture was taken, and the total broth was processed in the same way as for small-scale samples except that the first centrifugation step was omitted.