Phospholipids (PL) have gained the interest of academia and the food industry alike over the last two decades as nutraceuticals (commonly in infant formula and fitness nutrition) and as a functional ingredient, aiding in the instantization of powdered food products.  The most prevalent PL in dairy matrices are phosphatidylinositol (PI), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylcholine (PC), and sphingomyelin (SM).  Analysis of PL in published literature range from less expensive chromatography techniques like thin-layer chromatography to larger investments in instrumentation methods with varying detection techniques including fluorescence spectroscopy, evaporative light scattering, and mass spectrometry.  Methods from literature are often difficult to adopt as plug-and-play solutions to starting analysis of PL in industry, and typically require some degree of modification for acceptable results on the specific system configuration being used.  A rigorous, reproducible analytical method is required in an industry setting that allows for consistent analysis of food products at several location, in potentially more than one country.  The method generated in this collaboration between industry and academia has been fully validated for accuracy and reproducibility including intermediate precision within a group of industry scientists, resulting in ≤0.6% variation in total PL content measured compared to third party contracted analysis.  This method applies a modified Folch extraction procedure for sample preparation and clean-up followed by UPLC-ELSD analysis using HILIC chemistry on a silica normal phase column.  Briefly, powdered whey is homogenized in 125mM saline solution and extracted following the Folch method using 2:1 v/v chloroform-methanol.  After drying down the recovered lipids and reconstitution in extraction solvent the samples are analyzed on a Waters Acquity UPLC-ELSD system using a quaternary solvent system comprised of aqueous ammonium formate (200mM), isopropyl alcohol, methanol, and acetonitrile.  Spike recovery samples yielded reproducible results for all measured PL at &gt;75%.

Phospholipid quantitation in whey matrices by HILIC chemistry UPLC-ELSD:  Industry developed, validated, and applied method for determining phospholipid composition

2019 Fall National Meeting

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