High-performance liquid chromatography of adult human bronchoalveolar lavage: Assay for phospholipid lung profile

Abstract High-performance liquid chromatography has been used to separate pulmonary phospholipids from adult human bronchoalveolar lavage. A solvent system consisting of acetonitrile—water (80:20) as solvent A and pure acetonitrile as solvent B was used with a silica column (Bio-Sil HP 10) coupled to an Si-100 Polyol precolumn. A linear gradient from 87.5 to 25% of solvent B was found to separate all biologically relevant surfactant phospholipids in the following sequence and composition: phosphatidic acid (1.1%), phosphatidylglycerol (10.6%), phosphatidylinositol (9.9%), phosphatidylethanolamine (3.6%), phosphatidylserine (4.5%), phosphatidylcholine (60.8%), sphingomyelin (8.1%) and lysophosphatidylcholine (1.6%). These results were very similar to the phospholipid pattern obtained by two-dimensional thin-layer chromatography. It is concluded that high-performance liquid chromatography is a useful and rapid method for the separation of phospholipids in biological fluids containing pulmonary surfactant.