TY - JOUR
T1 - Optimized metabolite extraction from blood serum for 1H nuclear magnetic resonance spectroscopy
AU - Tiziani, Stefano
AU - Emwas, Abdul Hamid
AU - Lodi, Alessia
AU - Ludwig, Christian
AU - Bunce, Christopher M.
AU - Viant, Mark R.
AU - Günther, Ulrich L.
N1 - Funding Information:
We thank the EU for supporting S.T., A.E., and A.L. in the context of the MOTET Marie Curie project and the NERC for an Advanced Fellowship to M.R.V. (NER/J/S/2002/00618). This work was performed at the HWB-NMR facility in Birmingham which is supported by the Wellcome Trust. We are indebted to Chenomx Inc. for use of their NMR metabolomics software.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2008/6/1
Y1 - 2008/6/1
N2 - Blood serum is commonly used for clinical diagnostics because its protein composition bears a wealth of information about the health of an organism. More recently the analysis of the small molecule composition, the metabolome, has received increased attention because the metabolite composition is influenced by many diseases, by the administration of drugs and toxins, and by the diet and life style of an individual. When nuclear magnetic resonance spectroscopy is used as an analytical tool it is often preferable to remove catalytically active proteins, in particular for longer measurements, because metabolite concentrations are otherwise in constant flux. Here we have compared different protocols for the separation of proteins and metabolites, including precipitation methods and ultrafiltration. Whereas most extraction methods involving protein precipitation deplete some metabolites, ultrafiltration is superior in retaining metabolite concentrations and offers excellent reproducibility. We also describe a new method to recover the hydrophobic fraction for ultrafiltration with good reproducibility.
AB - Blood serum is commonly used for clinical diagnostics because its protein composition bears a wealth of information about the health of an organism. More recently the analysis of the small molecule composition, the metabolome, has received increased attention because the metabolite composition is influenced by many diseases, by the administration of drugs and toxins, and by the diet and life style of an individual. When nuclear magnetic resonance spectroscopy is used as an analytical tool it is often preferable to remove catalytically active proteins, in particular for longer measurements, because metabolite concentrations are otherwise in constant flux. Here we have compared different protocols for the separation of proteins and metabolites, including precipitation methods and ultrafiltration. Whereas most extraction methods involving protein precipitation deplete some metabolites, ultrafiltration is superior in retaining metabolite concentrations and offers excellent reproducibility. We also describe a new method to recover the hydrophobic fraction for ultrafiltration with good reproducibility.
UR - http://www.scopus.com/inward/record.url?scp=43049140446&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2008.01.037
DO - 10.1016/j.ab.2008.01.037
M3 - Journal articles
C2 - 18312846
AN - SCOPUS:43049140446
SN - 0003-2697
VL - 377
SP - 16
EP - 23
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -
