Evaluation of air oxidized PAPC: A multi laboratory study by LC-MS/MS


Oxidized LDL (oxLDL) has been shown to play a crucial role in the onset and development of cardiovascular disorders. The study of oxLDL, as an initiator of inflammatory cascades, led to the discovery of a variety of oxidized phospholipids (oxPLs) responsible for pro-inflammatory actions. Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (PAPC) is frequently used by the scientific community as a representative oxPL mixture to study the biological effects of oxidized lipids, due to the high abundance of PAPC in human tissues and the biological activities of oxidized arachidonic acids derivatives. Most studies focusing on oxPAPC effects rely on in-house prepared mixtures of oxidized species obtained by exposing PAPC to air oxidation. Here, we described a multi-laboratory evaluation of the compounds in oxPAPC by LC-MS/MS, focusing on the identification and relative quantification of the lipid peroxidation products (LPPs) formed. PAPC was air-oxidized in four laboratories using the same protocol for 0, 48, and 72 h. It was possible to identify 55 different LPPs with unique elemental composition and characterize different structural isomeric species within these. The study showed good intra-sample reproducibility and similar qualitative patterns of oxidation, as the most abundant LPPs were essentially the same between the four laboratories. However, there were substantial differences in the extent of oxidation, i.e. the amount of LPPs relative to unmodified PAPC, at specific time points. This shows the importance of characterizing air-oxidized PAPC preparations before using them for testing biological effects of oxidized lipids, and may explain some variability of effects reported in the literature.

Publication DOI: https://doi.org/10.1016/j.freeradbiomed.2019.06.013
Divisions: College of Health & Life Sciences
College of Health & Life Sciences > Aston Pharmacy School
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > Chronic and Communicable Conditions
Additional Information: © 2019 The Authors. Published by Elsevier Inc. CC BY-NC-ND. Funding: EU H2020 funded project MASSTRPLAN (Grant number 675132; to all authors except TM) and German Federal Ministry of Education and Research (BMBF) within the framework of the e:Med research and funding concept for SysMedOS project (to MF) are gratefully acknowledged. Thanks are due to University of Aveiro, Marine Lipidomics Laboratory, Fundação para a Ciência e a Tecnologia (FCT, MECPortugal), European Union, QREN, Programa Operacional Factores de Competitividade (COMPETE) and FEDER for the financial support to QOPNA (FCT UID/QUI/00062/2019) and CESAM (UID/AMB/50017/2019) research units; Portuguese Mass Spectrometry Network (LISBOA-01-0145-FEDER-402-022125), FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020.
Uncontrolled Keywords: Multi-laboratory study,Oxidized phospholipids,PAPC,Reverse phase chromatography,Tandem mass spectrometry,Biochemistry,Physiology (medical)
Publication ISSN: 1873-4596
Last Modified: 08 Jul 2024 07:51
Date Deposited: 19 Aug 2019 09:40
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Related URLs: https://linking ... 891584919303478 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2019-11-20
Published Online Date: 2019-06-15
Accepted Date: 2019-06-10
Authors: Ni, Zhixu
Sousa, Bebiana C. (ORCID Profile 0000-0002-1356-6789)
Colombo, Simone
Afonso, Catarina B. (ORCID Profile 0000-0003-4828-7146)
Melo, Tania
Pitt, Andrew R. (ORCID Profile 0000-0003-3619-6503)
Spickett, Corinne M. (ORCID Profile 0000-0003-4054-9279)
Domingues, Pedro
Domingues, M. Rosário
Fedorova, Maria
Criscuolo, Angela

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