title: Chromatography of acidic phospholipids on immobilized neomycin.
abstract: Columns of immobilized neomycin (reductively coupled to porous glass beads) were used to separate weakly acidic lipids as well as the polyphosphoinositides. All anionic lipids present in chloroform-methanol extracts, which had been washed first with acid and then with neutral salt solutions, were adsorbed. Phosphatidylserine and phosphatidic acid were eluted with chloroform-methanol-formic acid mixtures. Phosphatidylinositol and cardiolipin were eluted sequentially by very low concentrations (10-100 mM) of ammonium formate in chloroform-methanol-water. All three phosphoinositides were isolated from washed chloroform-methanol-KCl extracts of brain. Sulfatides eluted with the phosphatidylinositol. Phosphatidylinositol phosphate and phosphatidylinositol bisphosphate were eluted in turn by higher salt concentrations (0.4-1 M). The immobilized neomycin was used repeatedly but the capacity eventually declined. This loss was reversed by sodium borohydride reduction.
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002846:Chromatography, Affinity / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D008099:Liver / Q000032:analysis*; D009355:Neomycin; D010743:Phospholipids / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D010743:Phospholipids; D009355:Neomycin