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.
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.
Tuytten
Robin
R
Department of Chemistry, Nucleoside Research and Mass Spectrometry Unit and Center for Proteomics and Mass Spectrometry, University of Antwerp, Antwerp, Belgium.
Lemière
Filip
F
Van Dongen
Walter
W
Witters
Erwin
E
Esmans
Eddy L
EL
Newton
Russell P
RP
Dudley
Ed
E
D000818:Animals; D001897:Boronic Acids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003404:Creatinine / Q000652:urine; D006801:Humans; D009705:Nucleosides / Q000652:urine*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013997:Time Factors
D001897:Boronic Acids; D009705:Nucleosides; D003404:Creatinine