TY - JOUR
T1 - Global metabolic profiling using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry
AU - Qi, Yunpeng
AU - Song, Yunlong
AU - Gu, Haiwei
AU - Fan, Guorong
AU - Chai, Yifeng
PY - 2014/1/1
Y1 - 2014/1/1
N2 - Currently, liquid chromatography-mass spectrometry (LC-MS) is one of the most important analytical technologies for detecting hundreds of metabolites in the field of metabolomics. A recent advance in LC that has impacted metabolomics is the development of UPLC (ultra-performance liquid chromatography). In this chapter, we describe the analytical methodologies for the global metabolic profiling of serum, urine, and tissue samples using UPLC-Q-TOF (quadrupole-time-of-flight)-MS. Aqueous metabolites are extracted after adding methanol/acetonitrile/acetone and then analyzed by UPLC-MS under positive and/or negative ionization mode. With the aid of multivariate statistical analysis, separation between various groups can be observed in the score plots, and biomarkers are screened in the loading/ weight/VIP (variable importance in the projection) scatterplots. Furthermore, putative markers can be identified through comparison with the authentic standards based on tandem mass spectrometry (MS/MS) fragmentation pattern and LC retention. We expect that our protocol, with modifications if necessary, can be useful in many metabolomics studies and a wide range of research areas related to small molecules and LC-MS.
AB - Currently, liquid chromatography-mass spectrometry (LC-MS) is one of the most important analytical technologies for detecting hundreds of metabolites in the field of metabolomics. A recent advance in LC that has impacted metabolomics is the development of UPLC (ultra-performance liquid chromatography). In this chapter, we describe the analytical methodologies for the global metabolic profiling of serum, urine, and tissue samples using UPLC-Q-TOF (quadrupole-time-of-flight)-MS. Aqueous metabolites are extracted after adding methanol/acetonitrile/acetone and then analyzed by UPLC-MS under positive and/or negative ionization mode. With the aid of multivariate statistical analysis, separation between various groups can be observed in the score plots, and biomarkers are screened in the loading/ weight/VIP (variable importance in the projection) scatterplots. Furthermore, putative markers can be identified through comparison with the authentic standards based on tandem mass spectrometry (MS/MS) fragmentation pattern and LC retention. We expect that our protocol, with modifications if necessary, can be useful in many metabolomics studies and a wide range of research areas related to small molecules and LC-MS.
KW - Global metabolic profiling
KW - Liquid chromatography
KW - Mass spectrometry
KW - Metabolomics
KW - Quadrupole-time-of-flight
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U2 - 10.1007/978-1-4939-1258-2_2
DO - 10.1007/978-1-4939-1258-2_2
M3 - Article
C2 - 25270920
AN - SCOPUS:84921887344
SN - 1064-3745
VL - 1198
SP - 15
EP - 27
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -