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Profiling of polar metabolites in biological extracts using diamond hydride-based aqueous normal phase chromatography

Callahan, Damien L., De Souza, David, Bacic, Antony and Roessner, Ute 2009, Profiling of polar metabolites in biological extracts using diamond hydride-based aqueous normal phase chromatography, Journal of separation science, vol. 32, no. 13, pp. 2273-2280, doi: 10.1002/jssc.200900171.

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Title Profiling of polar metabolites in biological extracts using diamond hydride-based aqueous normal phase chromatography
Author(s) Callahan, Damien L.ORCID iD for Callahan, Damien L. orcid.org/0000-0002-6384-8717
De Souza, David
Bacic, Antony
Roessner, Ute
Journal name Journal of separation science
Volume number 32
Issue number 13
Start page 2273
End page 2280
Total pages 8
Publisher Wiley
Place of publication Londn, Eng.
Publication date 2009-07
ISSN 1615-9306
Keyword(s) Aqueous normal phase
ESI
Hydrophilic interaction chromatography
LC-MS
Metabolomics
Summary Highly polar metabolites, such as sugars and most amino acids are not retained by conventional RP LC columns. Without sufficient retention low concentration compounds are not detected due ion suppression and structural isomers are not resolved. In contrast, hydrophilic interaction chromatography (HILIC) and aqueous normal phase chromatography (ANP) retain compounds based on their hydrophilicity and therefore provides a means of separating highly polar compounds. Here, an ANP method based on the diamond hydride stationary phase is presented for profiling biological small molecules by LC. A rapid separation system based upon a fast gradient that delivers reproducible chromatography is presented. Approximately 1000 compounds were reproducibly detected in human urine samples and clear differences between these samples were identified. This chromatography was also applied to xylem fluid from soyabean (Glycine max) plants to which 400 compounds were detected. This method greatly increases the metabolite coverage over RP-only metabolite profiling in biological samples. We show that both forms of chromatography are necessary for untargeted comprehensive metabolite profiling and that the diamond hydride stationary phase provides a good option for polar metabolite analysis.
Language eng
DOI 10.1002/jssc.200900171
Field of Research 030108 Separation Science
0301 Analytical Chemistry
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2009, Wiley
Persistent URL http://hdl.handle.net/10536/DRO/DU:30083165

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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.