Gram‐scale preparative HPLC of phospholipids from soybean lecithins

A preparative procedure has been developed to isolate gram quantities of phospholipid classes from soybean lecithins. Various steps taken to accomplish the isolation are described—an analytical method with silica column and light scattering detector, alcohol fractionation of deoiled lecithins, and columns with increasing internal diameters but packed with the same stationary phase. The loading study showed that it was possible to inject 20 mg on a 100 × 8 mm RadialμPorasil colume. The separation was scaled up to a 25‐mm i.d. column and finally to a 50‐mm i.d. column. With the larger column, 2.1 g of phospholipids were separated. The collected fractions (phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid and phosphatidylcholine) were of high purity (>99%). The solvent consumption was 7.2 L (separation and column equilibration), and a minimum of 10 g of polar lipids can be separated daily.