Metabolomic profiling using Orbitrap Fourier transform mass spectrometry with hydrophilic interaction chromatography: a method with wide applicability to analysis of biomolecules.
It was shown that coupling hydrophilic interaction chromatography (HILIC) to Orbitrap Fourier transform mass spectrometery (FT-MS) provided an excellent tool for metabolic profiling, principally due to rapid elution of lipids in advance of most metabolites entering the mass spectrometer. We used in vitro cultivated procyclic forms of the protozoan parasite Trypanosoma brucei as a source of metabolites to test the performance of the HILIC column and the mass accuracy of MS. The mass accuracy achieved fell within 2 ppm for all the metabolites identified within samples. It was, for example, possible to identify the signature metabolite of the trypanosome, trypanothione, and also glutathione which were well retained by the HILIC column. By comparing trypanosomes grown in two different media we were able to clearly distinguish the samples in terms of the relative abundance of a number of metabolites using Sieve 1.1 software.
Kamleh
A
A
Strathclyde Institute for Pharmacy and Biomedical Sciences, University of Strathclyde, 27 Taylor Street, Glasgow G4 0NR, UK.
Barrett
M P
MP
Wildridge
D
D
Burchmore
R J S
RJ
Scheltema
R A
RA
Watson
D G
DG
D000818:Animals; D002845:Chromatography / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D046508:Culture Techniques; D005583:Fourier Analysis; D005947:Glucose / Q000737:chemistry; D005978:Glutathione / Q000031:analogs & derivatives / Q000737:chemistry; D008018:Life Cycle Stages; D013058:Mass Spectrometry / Q000379:methods*; D010743:Phospholipids / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D012984:Software; D013095:Spermidine / Q000031:analogs & derivatives / Q000737:chemistry; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*
D003470:Culture Media; D010743:Phospholipids; C044809:trypanothione; D011392:Proline; D005978:Glutathione; D005947:Glucose; D013095:Spermidine