A high-performance liquid chromatographic method for the analysis of lipids from lyophilized formulations

A simple, sensitive and rapid, high-performance liquid chromatography (HPLC) method has been developed for the simultaneous determination of cardiolipin, cholesterol and egg phosphatidylcholine from lyophilized lipid formulations. The proposed HPLC method utilizes a mu Porosil column (300x3.9 mm I.D., 10 mu m) and UV detection at 205 nm. A good resolution was observed with a mobile phase of water-methanol-acetonitrile (5:70:25) at a dow-rate of 2 ml/min. This method is capable of measuring levels of about 0.2-0.8 mg/ml of cardiolipin, 0.25-1.0 mg/ml of cholesterol and 0.8-3.2 mg/ml of egg phosphatidylcholine (EPC). The proposed analytical method has been validated according to USP compliance standards for accuracy, precision and reproducibility and thus may be employed for routine analysis of the lipid components from a typical placebo liposomal formulation.