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 co lumn (300x3.9 mm I.D., 10 mu m) and UV detection at 205 nm. A good res olution was observed with a mobile phase of water-methanol-acetonitril e (5:70:25) at a dow-rate of 2 ml/min. This method is capable of measu ring levels of about 0.2-0.8 mg/ml of cardiolipin, 0.25-1.0 mg/ml of c holesterol and 0.8-3.2 mg/ml of egg phosphatidylcholine (EPC). The pro posed 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.