Development of an on-line SPE-LC-ESI-MS method for urinary nucleosides: hyphenation of aprotic boronic acid chromatography with hydrophilic interaction LC-ESI-MS |
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| Authors: | Tuytten Robin Lemière Filip Van Dongen Walter Witters Erwin Esmans Eddy L Newton Russell P Dudley Ed |
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| Affiliation: | Department of Chemistry, Nucleoside Research and Mass Spectrometry Unit and Center for Proteomics and Mass Spectrometry, University of Antwerp, Antwerp, Belgium. |
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| Abstract: | The development of an on-line automated SPE-HPLC--ESI-MS method is described for targeted metabolomic analysis of urinary modified nucleoside levels. The setup comprises a boronate affinity column as a trapping device, a hydrophilic interaction chromatography (HILIC) separation and information-dependent MS detection modes. The system was optimized using standards and tested on biological samples, detecting a number of modified nucleosides. Other urinary biomarkers could also be analyzed by the system developed: for example, the urinary nucleobases were also available for analysis. A simultaneous creatinine-monitoring experiment was also demonstrated to be viable when utilizing the method, which is of benefit as creatinine is a urinary normalizing factor. |
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