Metabolome analysis via comprehensive two-dimensional liquid chromatography: identification of modified nucleosides from RNA metabolism

Analytical and Bioanalytical Chemistry
Lucas WillmannBernd Kammerer

Abstract

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 2...Continue Reading

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Citations

Dec 3, 2015·Bioanalysis·Dwight R Stoll
Jul 29, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Lucas WillmannBernd Kammerer
Feb 13, 2019·Future Medicinal Chemistry·Lauren EndresRebecca Rose
Jan 24, 2020·The Analyst·Marco Beccaria, Deirdre Cabooter
Jul 5, 2018·Journal of Otolaryngology - Head & Neck Surgery = Le Journal D'oto-rhino-laryngologie Et De Chirurgie Cervico-faciale·Philipp Opitz, Olf Herbarth
Jan 21, 2021·Molecular Cell·Akiko OgawaFan-Yan Wei
Jan 28, 2021·International Journal of Molecular Sciences·Faisal HayatMarie E Migaud
Mar 9, 2019·Journal of the American Society for Mass Spectrometry·Liqing HeXiang Zhang
Jun 25, 2021·World Journal of Gastrointestinal Oncology·Peng GaoYong Zhang
Jul 28, 2021·Annual Review of Analytical Chemistry·Stephanie L CollinsAndrew D Patterson
Dec 10, 2016·Analytical Chemistry·Dwight R Stoll, Peter W Carr

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