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Evaporate the solvent and add 5 ml methanolic KOH. Warm for 1 h at 80°C in a water or a sand bath. After cooling, extract the non-saponifiables with 2 washings of 5 ml diethyl ether. Add a few drops of phenolphthalein indicator to the lower phase and acidify with HCl (about 0.3 ml). Extract the fatty acids with 2 washings of 5 ml hexane. When short-chain fatty acids are present in unprecedented the lipid extract, it is necessary to unprecedented extract more unprecedented extensively with hexane (5 or 6 times). Do not evaporate too extensively the hexane phase (keep at a mild temperature) to prevent loss of these fatty acids. Fatty acids may be weighed, titrated to determine their neutralization equivalent or converted to methyl esters before fractionation or GLC analysis.. An alternative method for saponification has been proposed using a microwave-assisted treatment (Pineiro-Avila G et al.,
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