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Metabolome analysis via comprehensive two-dimensional liquid chromatography: identification of modified nucleosides from RNA metabolism
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  • 作者:Lucas Willmann ; Thalia Erbes ; Sonja Krieger…
  • 关键词:Modified nucleosides ; Metabolomics ; Two ; dimensional liquid chromatography ; 2D ; LC ; MS ; Comprehensive 2D ; LC ; Breast cancer
  • 刊名:Analytical and Bioanalytical Chemistry
  • 出版年:2015
  • 出版时间:May 2015
  • 年:2015
  • 卷:407
  • 期:13
  • 页码:3555-3566
  • 全文大小:2,013 KB
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  • 作者单位:Lucas Willmann (1) (3)
    Thalia Erbes (4)
    Sonja Krieger (2)
    Jens Trafkowski (2)
    Michael Rodamer (2)
    Bernd Kammerer (1)

    1. Center for Biological Systems Analysis ZBSA, Albert-Ludwigs-University Freiburg, Habsburgerstr. 49, 79104, Freiburg, Germany
    3. Institute of Biology II, Albert-Ludwigs-University Freiburg, Sch盲nzlestr. 1, 79104, Freiburg, Germany
    4. University Medical Center Freiburg, Hugstetter Stra脽e 55, 79106, Freiburg, Germany
    2. Agilent Technologies, Hewlett-Packard-Stra脽e 8, 76337, Waldbronn, Germany
  • 刊物类别:Chemistry and Materials Science
  • 刊物主题:Chemistry
    Analytical Chemistry
    Food Science
    Inorganic Chemistry
    Physical Chemistry
    Monitoring, Environmental Analysis and Environmental Ecotoxicology
  • 出版者:Springer Berlin / Heidelberg
  • ISSN:1618-2650
文摘
Modified nucleosides derived from the RNA metabolism constitute an important chemical class, which are discussed as potential biomarkers in the detection of mammalian breast cancer. Not only the variability of modifications, but also the complexity of biological matrices such as urinary samples poses challenges in the analysis of modified nucleosides. In the present work, a comprehensive two-dimensional liquid chromatography mass spectrometry (2D-LC-MS) approach for the analysis of modified nucleosides in biological samples was established. For prepurification of urinary samples and cell culture supernatants, we performed a cis-diol specific affinity chromatography using boronate-derivatized polyacrylamide gel. In order to establish a 2D-LC method, we tested numerous column combinations and chromatographic conditions. In order to determine the target compounds, we coupled the 2D-LC setup to a triple quadrupole mass spectrometer performing full scans, neutral loss scans, and multiple reaction monitoring (MRM). The combination of a Zorbax Eclipse Plus C18 column with a Zorbax Bonus-RP column was found to deliver a high degree of orthogonality and adequate separation. By application of 2D-LC-MS approaches, we were able to detect 28 target compounds from RNA metabolism and crosslinked pathways in urinary samples and 26 target compounds in cell culture supernatants, respectively. This is the first demonstration of the applicability and benefit of 2D-LC-MS for the targeted metabolome analysis of modified nucleosides and compounds from crosslinked pathways in different biological matrices.

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