In-Depth Lipidomic Analysis of Molecular Species of Triacylglycerides, Diacylglycerides, Glycerophospholipids, and Sphingolipids of Buttermilk by GC-MS/FID, HPLC-ELSD, and UPLC-QToF-MS

Abstract

Buttermilk, a byproduct of butter manufacturing, has gained considerable attention due to its high concentration of polar lipids as phospho- and sphingolipids from the milk fat globule membrane (MFGM). These polar lipids (PLs) are essential components of all cellular membranes and exert a variety of indispensable metabolic, neurological, and intracellular signaling processes. Despite its importance, there are few research studies that report a comprehensive characterization of the lipid molecular species of MFGM that could contribute to a better understanding of their putative healthful activities. In this study, procedures such as pressurized liquid extraction of polar and nonpolar lipids and their fractionation by flash chromatography have been carried out. The obtained fractions were submitted to an exhaustive characterization from a lipidomic point of view. The characterization includes new data about the identification and quantification of triacylglycerides (TAG), diacylglycerides (DAG), and phospho- and sphingolipids using different chromatographic techniques. The fatty acid profile was comparable to that of the milk fat but with a highly diverse composition of fatty acids. Molecular species have also been determined by using ultra-high performance liquid chromatography/quadruple-time-of-flight mass spectrometry (UPLC/QToF-MS). The TAG (16:0/16:0/6:0) and TAG (16:0/16:0/8:0) were the predominant saturated TAG species and TAG (14:0/18:1/16:0) and TAG (16:0/16:0/18:1) presented the highest content of monounsaturated TAG species. Furthermore; over 30 molecular species of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol (PI) could be identified within PL, with PC (16:0/18:1) being the most abundant species. Whereas C16:0 was found to be the preferred FA in TAGs, it was C18:1 in PLs. Several ganglioside species have also been characterized with d18:1 ceramide moiety and secondary acyl chains ranging from C20:0 to C26:1. This approach could broaden the applications of high-resolution mass spectrometry for a better understanding of the role of MFGM and its functionality.

Sources, Production, and Clinical Treatments of Milk Fat Globule Membrane for Infant Nutrition and Well-Being

“…This high protein content represents an interesting feature in the context of nutritional products such as IF or other dairy products. The detailed protein composition of MFGM-enriched ingredients is not systematically disclosed in technical data sheets; however, most the ingredients prepared from cream, beta serum, or buttermilk contain measurable amounts of gangliosides, MFGM proteins, cholesterol, sialic acid, IgG, and lactoferrin [ 41 , 42 , 43 , 44 , 45 ].…”

Milk fat globule membrane: An overview with particular emphasis on its nutritional and health benefits

“…In‐depth lipidomic analysis revealed the presence of more than 30 molecular species of PC, PE, PS and PI, of which PC (C16:0/C18:1) was the most abundant species in buttermilk (Castro‐Gómez et al . 2017). Several ganglioside species have also been characterised with d18:1 ceramide moiety and secondary acyl chains ranging from C20:0 to C26:1.…”

“…The composition and structure of MFGM became better understood with the advent of new tools and methods of analysis (Castro‐Gómez et al . 2017; Lopez et al . 2019; Wang et al, 2019; Calvo et al .…”

Metabolic Infrastructure of Pregnant Women With Trisomy 21 Fetuses; Metabolomic Analysis

“…2,5-Dioxopentanoate, a short-chain keto acid, is the main mediator of fatty aldehyde dehydrogenase which catalyzes the oxidation of long-chain aliphatic aldehydes to fatty acids. 1-Alkyl,2-acylglycerophosphoglycerol and 1-nanodecanoyl-2-3-sn-glycerol are cell membrane phospholipids, most probably acting as lipid messengers and regulators of cell growth and apoptosis [26].…”