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.