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Tandem LC columns for the simultaneous retention of polar and nonpolar molecules in comprehensive metabolomics analysis
Author(s) -
Chalcraft Kenneth R.,
McCarry Brian E.
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300779
Subject(s) - metabolomics , chemistry , tandem , chromatography , tandem mass spectrometry , chemical polarity , polar , molecule , mass spectrometry , materials science , organic chemistry , physics , astronomy , composite material
The tandem use of hydrophilic interaction LC columns with RP columns in series configuration has resulted in the retention of both polar and nonpolar components in complex biological samples (mouse serum) in a single analysis. This approach successfully coupled various columns with orthogonal separation characteristics, employed a single solvent gradient program compatible with the two columns and used ESI coupled to a TOF mass spectrometer for detection. Ion suppression, a common problem in ESI , was virtually eliminated for components eluting with apparent capacity factors >0.7. Retention time reproducibility with the tandem columns performed over three days with over 100 injections was comparable to that observed for single columns alone. This method was applied to the analysis of a pooled mouse serum sample and afforded highly reproducible data for up to 3000 mass spectral features. This approach was implemented with a conventional LC – MS system and should find broad applicability in the comprehensive analysis of complex mixtures containing a wide range of compound polarities.