Improvement in Resolution of Low Melting Point Triglyceride...

Published: Journal of Chromatographic Science, Volume 34, Number 7, July 1996, pp. 341-348.

Improvement in Resolution of Low Melting Point Triglyceride Molecular Species on Chilled Column Isocratic Liquid Chromatography
Tsugihiko Hirano and Koretaro Takahashi

An isocratic reversed-phase high-performance liquid chromatographic system is used to achieve the separation and improved resolution of individual triglyceride molecular species that are rich in icosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The triglycerides are enzymatically synthesized and contain EPA and DHA. Various column temperatures, from –20°C to 20°C, are used to aid in calculation of the changes in chromatographic parameters. It is found that when column temperature decreases, the selectivity values and retention rate values in the equation used to describe resolution exhibit an increasing; however, there is a critical column temperature that gives optimum resolution. For this reason, we concluded that to obtain the best resolution of a triglyceride molecular species with a low melting point, it is important to lower the column temperature near to the critical point.

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