Sackler School of
Graduate Biomedical Sciences

Lipid Extraction – Bligh and Dyer

For every 800 uL aqueous sample, add:

2 ml methanol, vortex
1 ml chloroform, vortex till 1 phase
1 ml chloroform, vortex
1 ml dw, vortex

Spin if necessary to separate phases.
Bottom fraction has lipids.

Pure B&D lower phase = chloroform/methanol (7.4:1)

Pure B&D upper phase = chloroform/methanol/dw (15:175:180)

Reference: Bligh and Dyer (1959)

Lipid Extraction – Folch

Measure volume sample.
Add 20 volumes chloroform/methanol (2:1). Vortex.
Add 4 volumes 0.9% NaCl. Vortex.
Spin in Accuspin if necessary to separate phases (about 2000 rpm, 10 min)
Bottom chloroform layer will have lipids. Wash with pure Folch upper.

Pure Folch upper = methanol/water/chloroform (94:96:4)

Pure Folch lower = methanol/water/chloroform (24:0.5:175)

Reference: Folch, Lees and Sloane Stanley (1957)
Modified September 2006