A Schlitt

A Zamyslowska

A Zhang

AC Schrimpe-Rutledge

B Zhou

CH Johnson

D Vuckovic

DG Sitnikov

E Rico

EJ Want

H Gika

J Godzien

K Skova

M Ciborowski

MC Cheung

RE Patterson

SJ Bruce

W Ebeling

X Li

Y Yang

Z Chen

AbstractPlasma untargeted metabolomics is a common method for evaluation of the mechanisms underlying human pathologies and identification of novel biomarkers. The plasma proteins provide the environment for transport of hydrophobic metabolites. The current sample preparation protocol relies on the immediate precipitation of proteins and thus leads to co-precipitation of a significant fraction of hydrophobic metabolites. Here we present a new simple procedure that overcomes the co-precipitation problem and improves metabolome coverage. Introducing an additional step preceding the protein precipitation, namely limited digestion with proteinase K, allows release of associated metabolites through the relaxation of the native proteins tertiary structure. The modified protocol allows clear detection of hydrophobic metabolites including fatty acids and phospholipids. Considering the potential involvement of the hydrophobic metabolites in human cardiovascular and cancer diseases, the method may constitute a novel approach in plasma untargeted metabolomics.</jats:p

Renata Wawrzyniak

Anna Kosnowska

Szymon Macioszek

Rafał Bartoszewski

Michał Jan Markuszewski

Crossref

Scientific Reports

New plasma preparation approach to enrich metabolome coverage in untargeted metabolomics: plasma protein bound hydrophobic metabolite release with proteinase K

New plasma preparation approach to enrich metabolome coverage in untargeted metabolomics: plasma protein bound hydrophobic metabolite release with proteinase K

Abstract

Similar works

Full text

Available Versions

Crossref