Quality control of liposomal lipids with special emphasis on peroxidation of phospholipids and cholesterol

The usefulness of various assays for the determination of phospholipid and cholesterol peroxidation in liposome formulations was studied on model liposomes prepared as small unilamellar vesicles (SUV) and multilamellar vesicles (MLV) from either native egg phosphatidylcholine (EPC), partially hydrogenated egg phosphatidylcholine (PHEPC) or fully hydrogenated egg phosphatidylcholine (HEPC) and cholesterol in 65/35 molar ratio at a total lipid concentration of 10 mumol/ml in phosphate buffered saline pH 7.2. Liposomes were incubated at 50 degrees C for a total of 3 months. Fatty acid and cholesterol peroxidation were monitored after 1, 2 and 3 months by quantitative measurement of fatty acids and cholesterol and as well as peroxidation products. Fatty acid peroxidation products malondialdehyde, lipidhydroperoxides, conjugated dienes, conjugated trienes were poor predictors of actual fatty acid loss. Among the cholesterol peroxidation products 7-hydroxy-cholesterols, 7-keto-cholesterol and 4-cholesten-3-one were measured quantitatively. Only the formation of 7-keto-cholesterol correlated well with cholesterol disappearance.