Separation and quantitation of water soluble cellular metabolites by hydrophilic interaction chromatography-tandem mass spectrometry.
A key unmet need in metabolomics is the ability to efficiently quantify a large number of known cellular metabolites. Here we present a liquid chromatography (LC)-electrospray ionization tandem mass spectrometry (ESI-MS/MS) method for reliable measurement of 141 metabolites, including components of central carbon, amino acid, and nucleotide metabolism. The selected LC approach, hydrophilic interaction chromatography with an amino column, effectively separates highly water soluble metabolites that fail to retain using standard reversed-phase chromatography. MS/MS detection is achieved by scanning through numerous selected reaction monitoring events on a triple quadrupole instrument. When applied to extracts of Escherichia coli grown in [12C]- versus [13C]glucose, the method reveals appropriate 12C- and 13C-peaks for 79 different metabolites.
Bajad
Sunil U
SU
Lewis-Sigler Institute for Integrative Genomics and Department of Chemistry, Princeton University, Princeton, NJ 08544, USA.
Lu
Wenyun
W
Kimball
Elizabeth H
EH
Yuan
Jie
J
Peterson
Celeste
C
Rabinowitz
Joshua D
JD
D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D003065:Coenzyme A / Q000032:analysis; D004926:Escherichia coli / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D009705:Nucleosides / Q000032:analysis; D009711:Nucleotides / Q000032:analysis; D015203:Reproducibility of Results; D012995:Solubility; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014867:Water / Q000737:chemistry*
D000596:Amino Acids; D002241:Carbohydrates; D009705:Nucleosides; D009711:Nucleotides; D014867:Water; D003065:Coenzyme A