A UPLC-MRM-MS method for comprehensive profiling of Amadori compound-modified phosphatidylethanolamines in human plasma
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RESEARCH PAPER
A UPLC-MRM-MS method for comprehensive profiling of Amadori compound-modified phosphatidylethanolamines in human plasma Xiaobo He 1,2 & Zhucui Li 1 & Qibin Zhang 1,3 Received: 30 April 2020 / Revised: 28 September 2020 / Accepted: 15 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Phosphatidylethanolamines (PEs) are targets of non-enzymatic glycation, a chemical process that occurs between glucose and primary amine-containing biomolecules. As the early-stage non-enzymatic glycation products of PE, Amadori-PEs are implicated in the pathogenesis of various diseases. However, only a few Amadori-PE molecular species have been identified so far; a comprehensive profiling of these glycated PE species is needed to establish their roles in disease pathology. Herein, based on our previous work using liquid chromatography-coupled neutral loss scanning and product ion scanning tandem mass spectrometry (LC-NLS-MS and LC-PIS-MS) in tandem, we extend identification of Amadori-PE to the low-abundance species, which is facilitated by using plasma lipids glycated in vitro. The confidence of identification is improved by high-resolution tandem mass spectrometry and chromatographic retention time regression. A LC-coupled multiple reaction monitoring mass spectrometry (LC-MRM-MS) assay is further developed for more sensitive quantitation of the Amadori compound-modified lipids. Using synthesized stable isotope-labeled Amadori lipids as internal standards, levels of 142 Amadori-PEs and 33 Amadori-LysoPEs are determined in the NIST human plasma standard reference material. These values may serve as an important reference for future investigations of Amadori-modified lipids in human diseases. Keywords Amadori compound . PE . LPE . LC-MRM-MS . Human plasma . NIST SRM-1950
Introduction Phosphatidylethanolamines (PEs), together with their partial hydrolysis products lysophosphatidylethanolamines (LPEs), are the second most abundant class of glycerophospholipids in human blood plasma [1]. Because of the primary amine in their ethanolamine head group, PEs and LPEs are targets of non-enzymatic glycation, also known as the Maillard reaction, in which the Supplementary Information The online version of this article (https:// doi.org/10.1007/s00216-020-03012-w) contains supplementary material, which is available to authorized users. * Qibin Zhang [email protected] 1
Center for Translational Biomedical Research, University of North Carolina at Greensboro, North Carolina Research Campus, Kannapolis, NC 28081, USA
2
Present address: National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA
3
Department of Chemistry & Biochemistry, University of North Carolina at Greensboro, Greensboro, NC 27412, USA
carbonyl group of reducing sugars reacts with the primary amines to initially form an unstable Schiff base and then a more stable Amadori product after rearrangement [2]. Although most of the literature points to the importance of protein glycation in
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