Solvent Systems - Lipid Migration

Chloroform:Methanol:Water (65:25:4)

Purpose: General separation of phospholipids by headgroup polarity. Notes: Phosphatidylserine, phosphatidylcholine, phosphatidylinositol, co-migrate in this solvent system. Phosphatidylglycerol and cardiolipin co-migrate in this solvent system.

Chloroform:Methanol:Ammonium Hydroxide (65:25:4)

Purpose: General separation of phospholipids by headgroup polarity and charge. Notes: Phosphatidylserine, phosphatidic acid, and phosphatidylinositol do not migrate above the origin in this solvent system. Phosphatidylglycerol and cardiolipin co-migrate in this solvent system.

Chloroform:Hexane:Methanol:Acetic Acid (50:30:10:5)

Purpose: Separation of cardiolipin and phosphatidylglycerol.

Toluene:Pyridine:Water (60:60:10)

Purpose: Separation of phosphatidylcholine, phosphatidyl(N,N-dimethylethanolamine), phosphatidyl(N- monomethylethanolamine), phosphatidylethanolamine.

Cyclohexane:Ethyl Acetate (3:2)

Purpose: Separation of neutral lipids (fatty acids, triglycerides, diglycerides, monoglycerides)

Toluene:Chloroform:Methanol (85:15:5)

Purpose: Separation of diglycerides (1,2 from 1,3 isomer).

Toluene:Methanol (7:3)

Purpose: Separation of ceramide from other sphingolipids. Notes: Rf of sphingolipids: Ceramide=0.57; Sphingosine=0.14; Sphingomyelin=0.04

Lipids i

Solvent System (Rf) [see chart for system] ii iii iv v vi

Triglycerides Free Fatty Acid Diglycerides 1,2 Diglycerides 1,3 Monoglycerides Phosphatidylethanolamine Phosphatidyl (Monomethylethanolamine) Cardiolipin Phosphatidylglycerol Phosphatidyl (Dimethylethanolamine) Phosphatidic Acid Phosphatidylinositol Phosphatidylcholine Phosphatidylserine Cerebrosides Ceramides Sphingosine Sphingomyelin Synthetic Cardiolipin Lyso-Phosphatidylglycerol Lyso-Phosphatidylethanolamine Monolysocardiolipin Lyso-Phosphatidic Acid Dilysocardiolipin 0.79 0.71 0.67 0.60 0.58 0.55 0.39 0.34 0.33 0.94 sf* 0.28 0.28 0.29 0.54 0.45 0.45 0.40 0.32 0.55 0.41 0.56 0.50 0.56 0.05 0.10 0.30 0.12 0.55 sf* 0.75 0.13 0.20 0.20 0.20 0.34 0.01 0.21 0.17 0.58 0.38 0.31 0.36 0.27 0.43 0.33

0.70 0.51 0.70

0.96 0.16 0.24 0.32

Lyso-Phosphatidylinositol Lyso-Phosphatidylcholine Lyso-Phosphatidylserine

i ii iii iv v vi

0.29 0.22 0.18

0.03 0.08 0.02

Ratio(V:V) 65:25:4 65:25:4 50:30:10:5 60:60:10 3:2 85:15:5

Solvent Systems Chloroform:Methanol:Water Chloroform:Methanol:Ammonium Hydroxide Chloroform:Hexane:Methanol:Acetic Acid Toluene:Pyridine:Water Cyclohexane:Ethyl Acetate Toluene:Chloroform:Methanol

Note:
Rf values represent relative migration only, whereas absolute values depend on various environmental parameters (e.g. temperature, humidity) which may vary depending on location. *sf - solvent front Avanti Polar Lipids, Inc. All rights reserved.Terms and ConditionsPrivacy PolicyComments