Deuterium Oxide Labeling for Global Omics Relative Quantification: Application to Lipidomics

Abstract

Partial metabolic deuterium oxide (D2O) combined with liquid chromatography-mass spectrometry (MS) has been utilized to quantify the in vivo turnover rate of an individual protein or lipid on a global scale. Herein, we developed a new global scale relative quantification method for lipids based on partial metabolic D2O labeling coupled with MS and named it “Deuterium Oxide Labeling for Global Omics Relative Quantification (DOLGOReQ). DOLGOReQ relies on subtle modification of isotope distributions of biomolecules induced by partial metabolic labeling of D2O. Fig. 1 displays the principle of DOLGOReQ. The accuracy, precision, and dynamic range of DOLGOReQ were evaluated with unlabled and D-labeled lipids from HeLa cells cultivated in normal and 5% D2O-enriched media, respectively. The number of detectable mass isotopomers and the extent of D-labeling were employed to filter out low quality quantification results using in-house software tailored for high-throughput data analysis of DOLGOReQ. DOLGOReQ demonstrated effective D-labeling and quantification of various lipid classes and provided two orders of magnitude of effective quantification dynamic range. Finally, relative quantification of HeLa lipids grown in normoxic and hypoxic conditions was performed with DOLGOReQ as a proof-of-concept experiment.