High-Purity Separation of Phospholipids by Preparative High-Performance Liquid Chromatography

초록

Soybean lecithin (phospholipids), mainly phosphatidylethanolamine(PE), phosphatidyl-inositol(PI), and phosphatidylcholine(PC), is a commercial by-product of vegetable oil production. Phospholipids are functioning phosphatides in cell membranes and important substances in biomedical, pharmaceutical and nutrition research. They are widely based in the food and cosmetic industries, as well as industrial manufacturing. In recent years a considerable increase in the application of gradient elution in more complex analytical problems has been observed. The purpose of this work was to investigate the effects of particle size in a chromatographic column, and to obtain the optimum mobile phase with the particle sizes for preparative work. Normal-phase HPLC was applied. The ternary mobile phase of hexane, methanol, and IPA were used as mobile phases. With the commercially available Nova-Pack column (4mm packing size, 3.9mm i.d., 15cm length), the gradient condition and mobile phase composition was that hexane was linearly decreased from 85 to 0 vol.% in two steps and IPA and methanol were also linearly increased 10 to 30 and 5 to 70 vol.% in two steps, respectively during the total run time of 25 min at the mobile phase flow rate of 1.5 ml/min. The three preparative packing sizes experimented were 15, 5/20, and 25/40 mm. The resolutions between PE and PI, PI and PC with 15 mm packings filled in the same column size was better over the other two preparative packings. A slight change in mobile phase composition was applied to increase the resolution at the same mobile phase flow rate.