Analysis of Simple Lipids and Related Compounds Using a Single HPSEC Column

This work summarizes and discusses the experiences of using a single size exclusion (100-Å Phenogel) column for the separation of simple lipids and related compounds by HPSEC. Generally, multiple (2 to 4) columns connected in series are used to separate these compounds which takes a long analysis time and is costly for the columns. It is almost 30 years since a single Phenogel column has been used to analyze the simple lipids in transmethylation of palm oil first published. The attempts to offer an alternative method for the separations of lipid compounds in a single Phenogel column using a modified mobile phase are successful during the last ten years. A small amount of acetic acid could be used as a modifier to suppress peak tailing and improve the separation of the lipids containing polar hydroxyl and carboxyl groups such as diglyceride, monoglyceride, and free fatty acid. A mixture of good and poor swelling solvents could be used to control swelling of the ST-DVB and change the elution strength of the mobile phase and was used to separate the lipids of similar molecular weight such as the wax ester and triglyceride and vitamin E isomers.