title: Development of a direct in-matrix extraction (DIME) protocol for MALDI-TOF-MS detection of glycated phospholipids in heat-treated food samples.
abstract: In foodstuffs, one of the main factors inducing modifications in phospholipids (PLs) structure is the heat treatment. Among PLs, only phosphatidylethanolamines and phosphatidylserines, due to their free amino group, can be involved in Maillard reaction and can form adducts with reducing sugars, besides other by-products called advanced glycation end-products. To date, glycated lipid products are less characterized in comparison to proteins. The aim of this work was to develop a novel, rapid and sensitive extraction protocol for the detection and characterization of modified PLs (glycated and oxidized) by means of matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). At first, to investigate the formation of glycated and/or short chain by-products in different classes of PLs, representative standards were heated with or without sugar (lactose or glucose) and subjected to traditional lipid extraction methods as Bligh and Dyer and to the novel direct in matrix extraction (DIME) using 1,8-bis(dimethylamino)naphthalene as preconcentrating matrix. MALDI-MS analysis in negative ion mode allowed detecting glycation and oxidation products both on fatty acid and glucose moieties. Then, the procedure was successfully applied to different heat-treated and powdered samples (milk powders, pasteurized milk, ultra-high-temperature milk and soy flour) for the detection of modified PLs in complex foods. The currently developed DIME protocol could be a powerful tool for understanding lipid glycation also in biological samples.
authors.affiliation: Università degli Studi di Bari Aldo Moro, Dipartimento di Chimica, Via Orabona 4, 70126, Bari, Italy.
authors.forename: Cosima D
authors.initials: CD
authors.lastname: Calvano
authors.forename: Cristina
authors.initials: C
authors.lastname: De Ceglie
authors.forename: Carlo G
authors.initials: CG
authors.lastname: Zambonin
mesh_terms: D000818:Animals; D005591:Chemical Fractionation; D005504:Food Analysis / Q000379:methods*; D005511:Food Handling; D006031:Glycosylation; D008892:Milk / Q000737:chemistry; D010743:Phospholipids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D010743:Phospholipids
keywords: MALDI-MS; Maillard reaction; glycated lipids; heated food; proton sponge