Separations of Major Soybean Phospholipids on β-Cyclodextrinbonded Silica

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Abstract
The four major phospholipids (PL) phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidic acid (PA) found in soybean oil were separated by normal-phase high-performance liquid chromatography (HPLC) on β-cyclodextrin-bonded silica (CDS) with UV detection. Adequate base-line separations of the PL components were achieved by isocratic elution with mobile phases containing hexane, isopropanol, ethanol and water/tetramethylammonium phosphate (TMAP). The presence of TMAP in the mobile phases was critical to improve component resolution and enhance peak symmetry. Analyte retention and component separations were dramatically influenced by a small change in mobile phase compositions. Under the HPLC conditions employed, the elution order appeared to follow the order of increasing PL polarity with increasing retention times (PE > PC > PI > PA). The HPLC method was used in the qualitative analyses of selected commercial lecithin samples. Potential applicability of the HPLC-UV-CDS technique for the quantitative analysis of several crude oil samples derived from genetically modified soybean oil is demonstrated.

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