A method of isolation and complete separation of lipids from natural sources into classes and species is reported which combines our previously published techniques with new techniques described in this article for the first time. Pigments are separated from crude total lipid extracts with two successive TLC systems: a) petroleum ether/benzene/glacial acetic acid (30:70:2) and b) acetone/methanol/water (40:20:1). Pigment-free total lipids are separated on a silicic acid column into neutral, glyco- and phospholipids. Neutral, glyco- and phospholipids are separated into classes and species by suitable HPLC methods.