TY - JOUR
T1 - Evaluation of coupling reversed phase, aqueous normal phase, and hydrophilic interaction liquid chromatography with orbitrap mass spectrometry for metabolomic studies of human urine
AU - Zhang, Tong
AU - Creek, Darren John
AU - Barrett, Michael P
AU - Blackburn, Gavin
AU - Watson, David G
PY - 2012
Y1 - 2012
N2 - In this study, we assessed three liquid chromatographic platforms: reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction (HILIC) for the analysis of polar metabolite standard mixtures and for their coverage of urinary metabolites. The two zwitterionic HILIC columns showed high-quality chromatographic performance for metabolite standards, improved separation for isomers, and the greatest coverage of polar metabolites in urine. In contrast, on the reversed phase column, most metabolites eluted very rapidly with little or no separation. Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic platform, approximately 970 metabolite signals with repeatable peak areas (relative standard deviation (RSD) = 25 ) could be putatively identified in human urine, by elemental composition assignment within a 3 ppm mass error. The ability of the methodology for the verification of nonmolecular ions, which arise from adduct formation, and the possibility of distinguishing isomers could also be demonstrated. Careful examination of the raw data and the use of masses for predicted metabolites produced an extension of the metabolite list for human urine.
AB - In this study, we assessed three liquid chromatographic platforms: reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction (HILIC) for the analysis of polar metabolite standard mixtures and for their coverage of urinary metabolites. The two zwitterionic HILIC columns showed high-quality chromatographic performance for metabolite standards, improved separation for isomers, and the greatest coverage of polar metabolites in urine. In contrast, on the reversed phase column, most metabolites eluted very rapidly with little or no separation. Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic platform, approximately 970 metabolite signals with repeatable peak areas (relative standard deviation (RSD) = 25 ) could be putatively identified in human urine, by elemental composition assignment within a 3 ppm mass error. The ability of the methodology for the verification of nonmolecular ions, which arise from adduct formation, and the possibility of distinguishing isomers could also be demonstrated. Careful examination of the raw data and the use of masses for predicted metabolites produced an extension of the metabolite list for human urine.
UR - http://pubs.acs.org.ezproxy.lib.monash.edu.au/doi/pdf/10.1021/ac2030738
U2 - 10.1021/ac2030738
DO - 10.1021/ac2030738
M3 - Article
SN - 0003-2700
VL - 84
SP - 1994
EP - 2001
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 4
ER -