Analytical and Bioanalytical Chemistry

, Volume 407, Issue 17, pp 5161–5173 | Cite as

Identification of carbonylated lipids from different phospholipid classes by shotgun and LC-MS lipidomics

  • Zhixu Ni
  • Ivana Milic
  • Maria FedorovaEmail author
Research Paper
Part of the following topical collections:
  1. Lipidomics


Oxidized lipids play a significant role in the pathogenesis of numerous oxidative stress-related human disorders, such as atherosclerosis, obesity, inflammation, and autoimmune diseases. Lipid peroxidation, induced by reactive oxygen and nitrogen species, yields a high variety of modified lipids. Among them, carbonylated lipid peroxidation products (oxoLPP), formed by oxidation of the fatty acid moiety yielding aldehydes or ketones (carbonyl groups), are electrophilic compounds that are able to modify nucleophilic substrates like proteins, nucleic acid, and aminophospholipids. Some carbonylated phosphatidylcholines possess even pro-inflammatory activities. However, little is known about oxoLPP derived from other phospholipid (PL) classes. Here, we present a new analytical strategy based on the mass spectrometry (MS) of PL-oxoLPP derivatized with 7-(diethylamino)coumarin-3-carbohydrazide (CHH). Shotgun MS revealed many oxoLPP derived from in vitro oxidized glycerophosphatidylglycerols (PG, 31), glycerophosphatidylcholine (PC, 23), glycerophosphatidylethanolamine (PE, 34), glycerophosphatidylserines (PS, 7), glycerophosphatidic acids (PA, 17), and phosphatidylinositiolphosphates (PIP, 6) vesicles. This data were used to optimize LipidXplorer-assisted identification, and a python-based post-processing script was developed to increase both throughput and accuracy. When applied to full lipid extracts from rat primary cardiomyocytes treated with peroxynitrite donor SIN-1, ten PL-bound oxoLPP were unambiguously identified by LC-MS, including two PC-, two PE-, one PG-, two PS-, and three PA-derived species. Some of the well-known carbonylated PC were detected, while most PL-oxoLPP were shown for the first time.

Graphical Abstract

Overview of analytical and bioinformatics approach for detection and identification of carbonylated phospholipids.


7-(Diethylamino)coumarin-3-carbohydrazide Carbonylated lipids LC-MS Lipid peroxidation products LipidXplorer Shotgun lipidomics 



Arachidonic acid




Collision-induced dissociation




Double bond equivalents


Data-dependent acquisition


Docosahexaenoic acid




Fatty acid


Gas phase fractionation


Head group




Linoleic acid


Liquid chromatography coupled to mass spectrometry


Lipid peroxidation products


Molecular fragmentation query language


Mass spectrometry


Tandem mass spectrometry


Methyl-tert-butyl ether


Normalized collision energy


Oleic acid


Oxygen addition products


Oxygen cleavage products


Carbonylated lipid peroxidation products


Oxidized phosphatidylcholine


Glycerophosphatidic acids






















1-Palmitoyl-2-linoleoyl-glycerophosphatidic acid










1-Palmitoyl-2-oleoyl-glycerophosphatidic acid














Polyunsaturated fatty acid


Reactive nitrogen species


Reactive oxygen species


Reversed phase chromatography











The authors are thankful to Prof. Ralf Hoffmann (Institute of Bioanalytical Chemistry, University of Leipzig) for providing access to his laboratory and mass spectrometers. Financial support from European Regional Development Fund (ERDF, European Union and Free State Saxony; 100146238 and 100121468 to M.F) and a stipend to I.M. provided by Universität Leipzig are gratefully acknowledged.

Supplementary material

216_2015_8536_MOESM1_ESM.pdf (475 kb)
ESM 1 (PDF 474 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Institute of Bioanalytical Chemistry, Faculty of Chemistry and MineralogyUniversität LeipzigLeipzigGermany
  2. 2.Center for Biotechnology and BiomedicineUniversität LeipzigLeipzigGermany

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