Quantitative analysis of pulmonary surfactant phospholipids by high-performance liquid chromatography and light-scattering detection.

An improved high-performance liquid chromatographic method for the separation and quantitation of nine phospholipid classes is described. It is based on normal-phase chromatography with silica gel as stationary phase and a binary gradient with mixtures of chloroform, methanol and water as mobile phase. The response of the evaporative light-scattering detector was non-linear. Peak areas were proportional to the power 1.7 of the masses. Phospholipids in lung lavage samples were enriched by liquid extraction prior to HPLC analysis. The described method is a rapid and accurate procedure for the quantitative analysis of phospholipid classes in biological samples.