Article (Scientific journals)
An Easy, Convenient Cell and Tissue Extraction Protocol for Nuclear Magnetic Resonance Metabolomics.
Matheus, Nicolas; Hansen, Sylvain; Rozet, Eric et al.
2014In Phytochemical Analysis, 25, p. 342-349
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Keywords :
NMR; cells; metabolite extraction; metabolomics; sample preparation; sonication; tissues
Abstract :
[en] INTRODUCTION: As a complement to the classic metabolomics biofluid studies, the visualisation of the metabolites contained in cells or tissues could be a very powerful tool to understand how the local metabolism and biochemical pathways could be affected by external or internal stimuli or pathologies. Therefore, extraction and/or lysis is necessary to obtain samples adapted for use with the current analytical tools (liquid NMR and MS). These extraction or lysis work-ups are often the most labour-intensive and rate-limiting steps in metabolomics, as they require accuracy and repeatability as well as robustness. Many of the procedures described in the literature appear to be very time-consuming and not easily amenable to automation. OBJECTIVE: To find a fast, simplified procedure that allows release of the metabolites from cells and tissues in a way that is compatible with NMR analysis. METHODS: We assessed the use of sonication to disrupt cell membranes or tissue structures. Both a vibrating probe and an automated bath sonicator were explored. RESULTS: The application of sonication as the disruption procedure led to reproducible NMR spectral data compatible with metabolomics studies. This method requires only a small biological tissue or cell sample, and a rapid, reduced work-up was applied before analysis. The spectral patterns obtained are comparable with previous, well-described extraction protocols. CONCLUSION: The rapidity and the simplicity of this approach could represent a suitable alternative to the other protocols. Additionally, this approach could be favourable for high- throughput applications in intracellular and intratissular metabolite measurements. Copyright (c) 2014 John Wiley & Sons, Ltd.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Matheus, Nicolas ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > GIGA-R : Labo de recherche sur les métastases
Hansen, Sylvain ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biologie cellulaire et moléculaire appliquée à l'homme
Rozet, Eric ;  Université de Liège - ULiège > Département de pharmacie > Chimie analytique
Peixoto, Paul ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > GIGA-R : Labo de recherche sur les métastases
Maquoi, Erik  ;  Université de Liège - ULiège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
LAMBERT, Vincent  ;  Centre Hospitalier Universitaire de Liège - CHU > Ophtalmologie
Noël, Agnès ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biologie cellulaire et moléculaire appliquée à l'homme
Frederich, Michel  ;  Université de Liège - ULiège > Département de pharmacie > Pharmacognosie
Mottet, Denis ;  Université de Liège - ULiège > Département des sciences de la vie > Génétique et biologie moléculaires animales
De Tullio, Pascal ;  Université de Liège - ULiège > Département de pharmacie > Chimie pharmaceutique
Language :
English
Title :
An Easy, Convenient Cell and Tissue Extraction Protocol for Nuclear Magnetic Resonance Metabolomics.
Publication date :
January 2014
Journal title :
Phytochemical Analysis
ISSN :
0958-0344
eISSN :
1099-1565
Publisher :
Wiley
Volume :
25
Pages :
342-349
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2014 John Wiley & Sons, Ltd.
Available on ORBi :
since 24 January 2014

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