Semi-Preparative HPLC Fractionation of Consumer Fish Oil (Triacylglycerols)

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Abstract
A reversed-phase high performance liquid chromatographic method was developed for the semi-preparative fractionation of fish oil-derived triacylglycerols containing the esterified omega-3 fatty acids all-cis-5,8,11,14,17-eicosapentaenoic acid [EPA] and all-cis-4,7,10,13,17,20-docosahexaenoic acid [DHA]. Analytical separation conditions, such as mobile phase composition and flow rate could be directly applied to the semi-preparative mode, which was further optimized. Separation of triacylglycerol fractions was obtained in 15 minutes using flow rates of 3.0 ml/min with a mobile phase of acetone/acetonitrile (65:35, v/v). 250-mg samples of the fish oil were fractionated and multi-milligram quantities of triacylglycerols were separated, which were 65% enriched in esterified EPA and DHA; a production rate of 500 mg/hr of this enriched fraction was obtained.
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