Abstract
We have developed a high performance liquid chromatography (HPLC) method to separate lecithin from other phospholipid classes and to obtain lecithin from biologic materials. The separation was performed on a preparative 10-micron Spherisorb column with an optimized solvent system consisting of the following components: acetonitrile, isopropanol, methanol, water, and trifluoroacetic acid. The advantages of this method are the use of an isocratic solvent system limited to about 30 min and the very good separation of the phosphatidyl-choline fraction from the sphingomyelin fraction. Furthermore, the HPLC method has a better recovery rate than the thin-layer chromatography method, and it can be run under automatic control.
Highlights
Thin-layer chromatography (TLC) on silica gel is used widely for separation and identification of lipids extracted from biological samples [1,2,3,4]
Our special interest was focused on phosphatidylcholine as a critical component of lung surfactant phospholipids. We developed this high performance liquid chromatography (HPLC) method for the preparative separation of lecithin from all other phospholipid classes
Quantitation of lecithin species was achieved by gas-liquid chromatography (GLC) using an internal standard.Recovery was assessed by determining the inorganic phosphorus content of HPLC fractions relative to the amount in injected standards
Summary
Thin-layer chromatography (TLC) on silica gel is used widely for separation and identification of lipids extracted from biological samples [1,2,3,4]. Numerous high performance liquid chromatography (HPLC) methods for the separation of phospholipids have been published [5,6,7,8,9,10,11,12,13]. The use of HPLC for the analysis of phospholipids present in biological samples is still limited. We developed this HPLC method for the preparative separation of lecithin from all other phospholipid classes.
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