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Saponification of the lipid extracts

HA Hadil Alaydi
PD Peter Downey
MM Michelle McKeon-Bennett
TB Tanya Beletskaya
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All solvent and SFE-CO2 extracts were completely dried under nitrogen gas and saponified by adding to each extract 1500 μl of saponification solution (2.5 M KOH in methanol, 1:4 v/v), prior to being vortexed and incubated at 72 ∘C for 15 min. Following incubation, 225 μl of formic acid was added, 1725 μl of chloroform and 375 μl of milli-Q water. The final mixture was vortexed and centrifuged for 5 min at 3000 rpm where two layers formed. The organic layer containing free fatty acid and polyprenol was transferred to a clean vial, dried and stored at − 20 ∘C for further analysis 30.

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