PuSH - Publication Server of Helmholtz Zentrum München

Blume, B. ; Witting, M. ; Schmitt-Kopplin, P. ; Michalke, B.

Novel extraction method for combined lipid and metal speciation from Caenorhabditis elegans with focus on iron redox status and lipid profiling.

Front. Chem. 9:788094 (2021)
Publ. Version/Full Text DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Parkinson´s disease progression is linked to iron redox status homeostasis via reactive oxygen species (ROS) formation, and lipids are the primary targets of ROS. The determination of iron redox status in vivo is challenging and requires specific extraction methods, which are so far tedious and very time-consuming. We demonstrated a novel, faster, and less laborious extraction method using the chelator ethylene glycol l-bis(β-aminoethyl ether)-N,N,N',N'-tetra acetic acid (EGTA) as a stabilizing agent and synthetic quartz beads for homogenization under an argon atmosphere. Additionally, we combined the metal extraction with a well-established lipid extraction protocol using methyl-tert-butyl ether (MTBE) to avoid the problems of lipid precipitation in frozen samples and to determine lipid profiles and metal species from the same batch. The nonextractable matrix, such as the debris, is removed by centrifugation and digested to determine the total metal content of the sample as well. Lipid profiling using RP-LC-MS demonstrated high accordance of the modified extraction method to the reference method, and the organic solvent does not affect the iron redox status equilibrium. Furthermore, rigorous testing demonstrated the stability of the iron redox status equilibrium during the extraction process, secured by complexation, inert atmosphere, fast preparation, and immediately deep frozen extracts.
Altmetric
Additional Metrics?
[➜Log in]
Edit extra informations Login
Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords C. Elegans Model ; Extraction ; Iron (ii) ; Iron (iii) ; Lipidomics ; Metal Speciation; Quantitative-analysis; Parkinsons-disease; Chromatography; Mechanisms; Capillary; Copper; Model
ISSN (print) / ISBN 2296-2646
e-ISSN 2296-2646
Journal Frontiers in Chemistry
Quellenangaben Volume: 9, Issue: , Pages: , Article Number: 788094 Supplement: ,
Publisher Frontiers
Publishing Place Lausanne
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Research Unit Analytical BioGeoChemistry (BGC)
CF Metabolomics & Proteomics (CF-MPC)