Why do not just use the classical Folch protocol: chloroform: methanol 2:1 v/v containing 25 mg/L of BHT? And may I ask you, what are you going to do with extract further? Is it prepared for separation or just for fatty acids analysis of total lipids?

Agree, Folch is the classical and tested method for total lipid extraction from tissue samples (

Folch, J., Lees, M., and Sloane-Standley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipids from Animal Tissues, J. Biol. Chem. 226, 497–509.)

I want to quantify and analyze fatty acids. We are having trouble breaking yeast cells. Roman, what is BHT? Monica

BHT = butylated hydroxytoluene, a commonly used antioxidant to prevent lipid autooxidation during extraction and sample processing.

You can also use Bleigh and Dyer method for extraction and purification of lipids, but I agree that Folch, even if it takes more time is more efficient. Then, to metilate and separate the methil esters we have different protocols according to how you are going to quantify the FA,

Well, if you are not going to fractionate lipid extracts why you do not just use saponification of whole cells in the presence of inner standard with subsequent extraction of free fatty acids and their methanolysis?

I think you could go simply: use the methanolysis of whole cells as it was done by J. Ohlrogge in one of his paper :)

From my experience, chloroform:methanol alone is not sufficient for yeast. You have to break the cell wall to get efficient extraction. We did bead beating in a 1.5mL tube followed by folch extraction and could get complete extraction.