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A tandem liquid chromatography–mass spectrometry (LC–MS) method for profiling small molecules in complex samples

Pyke, James S., Callahan, Damien L., Kanojia, Komal, Bowne, Jairus, Sahani, Sheena, Tull, Dedreia, Bacic, Antony, McConville, Malcolm J. and Roessner, Ute 2015, A tandem liquid chromatography–mass spectrometry (LC–MS) method for profiling small molecules in complex samples, Metabolomics, vol. 11, no. 6, pp. 1552-1562, doi: 10.1007/s11306-015-0806-7.

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Title A tandem liquid chromatography–mass spectrometry (LC–MS) method for profiling small molecules in complex samples
Author(s) Pyke, James S.
Callahan, Damien L.ORCID iD for Callahan, Damien L. orcid.org/0000-0002-6384-8717
Kanojia, Komal
Bowne, Jairus
Sahani, Sheena
Tull, Dedreia
Bacic, Antony
McConville, Malcolm J.
Roessner, Ute
Journal name Metabolomics
Volume number 11
Issue number 6
Start page 1552
End page 1562
Total pages 11
Publisher Springer
Place of publication Berlin, Germany
Publication date 2015-12
ISSN 1573-3882
1573-3890
Keyword(s) Reversed phase
Aqueous normal phase
Mass spectrometry
Metabolomics
Tandem liquid chromatography
Summary Liquid chromatography–mass spectrometry (LC–MS) methods using either aqueous normal phase (ANP) or reversed phase (RP) columns are routinely used in small molecule or metabolomic analyses. These stationary phases enable chromatographic fractionation of polar and non-polar compounds, respectively. The application of a single chromatographic stationary phase to a complex biological extract results in a significant proportion of compounds which elute in the non-retained fraction, where they are poorly detected because of a combination of ion suppression and the co-elution of isomeric compounds. Thus coverage of both polar and non-polar components of the metabolome generally involves multiple analyses of the same sample, increasing the analysis time and complexity. In this study we describe a novel tandem in-line LC–MS method, in which compounds from one injection are sequentially separated in a single run on both ANP and RP LC-columns. This method is simple, robust, and enables the use of independent gradients customized for both RP and ANP columns. The MS signal is acquired in a single chromatogram which reduces instrument time and operator and data analysis errors. This method has been used to analyze a range of biological extracts, from plant and animal tissues, human serum and urine, microbial cell and culture supernatants. Optimized sample preparation protocols are described for this method as well as a library containing the retention times and accurate masses of 127 compounds.
Language eng
DOI 10.1007/s11306-015-0806-7
Field of Research 030101 Analytical Spectrometry
0301 Analytical Chemistry
1103 Clinical Sciences
0601 Biochemistry And Cell Biology
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2015, Wiley
Persistent URL http://hdl.handle.net/10536/DRO/DU:30081943

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