Development of quantitation method for glycated aminophospholipids at the molecular species level in powdered milk and powdered buttermilk

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Abstract
The Maillard reaction is a nonenzymatic glycation reaction between a reducing sugar and a free amino group, known to naturally occur during heat processing of food. In this study, we especially focused on phosphatidylethanolamine (PE)-linked Amadori products (Amadori-PE) in powdered milk, since the analysis of these products at the molecular species level has not yet been evaluated. Analysis of Amadori-PE was conducted by using liquid chromatography-tandem mass spectrometry in three different modes. The main Amadori-PE species in a powdered milk sample were first identified as 34:1, 36:1, 36:2 and 36:3 in the total ion current mode. Additionally, by using the characteristic product ions observed in the presence of sodium, we quantified the main Amadori-PE species in the multiple reaction monitoring mode, and evaluated their total concentrations in the precursor ion scan (PIS) mode for the first time. Powdered milk contained much Amadori-PE with concentrations ranging from 4.3 to 8239 mg/100 g, quantified by the PIS mode. The newly developed methods represent powerful tools for detailed analysis of glycated lipids including Amadori-PE in powdered milk, which may further be applied to research relating to infant food and nutrition.

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