Sample Preparation

All samples (50–100 mg) were extracted in 1.2 mL methanol/chloroform (1:1) for 15 min using vortex mixing. Extracts were then centrifuged to remove solids,

and 1.0 mL of extract was added to 4 mL of isopropanol/ water (3:2) and 1 mL of 5 M KOH. All liquid oil samples (50 μL aliquot) were dissolved/dispersed in 5 mL isopropanol/water (3:2) and 1 mL of 5 M KOH.

All samples were heated in an 80 °C water bath for 1 h with occasional stirring. After samples were cooled, a 500 μL aliquot was removed and 25 μL of formic acid

was added to neutralize the sample.

Try it

gudluck

:)

This paper may help you out.

Thank you Robin ji. As per your answer, in the final step neutralization with any acid is acceptable? is there any specific reason for choosing formic acid?

Yes you can do. Try this method.

When KOH is added,soap is created (sodium salt of fatty acid), which is soluble in water