Development of a purification procedure for the isolation of nucleosides from urine prior to mass spectrometric analysis.
A chromatographic separation of nucleosides from urine has been developed in order to facilitate their mass spectrometric analysis for clinical diagnosis. A number of chromatographic resins were studied in order to develop an effective and efficient purification procedure. The optimized sequential protocol comprises a centrifugation, acidification and neutralization step, followed by application of an affinity chromatographic column and finally further separation on an acidic cation exchange column and a basic anion exchanger. This scheme shows effective clean-up of a standard radiolabelled nucleoside with a recovery of 92.5%, and recovery of nucleosides added to urine samples before extraction showed recoveries of 72-82%.
Dudley
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Biochemistry Research Group, School of Biological Sciences, University of Wales Swansea, UK.
El-Shakawi
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Games
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Newton
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RP
D000180:Acrylic Resins / Q000737:chemistry; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003911:Dextrans / Q000737:chemistry; D005782:Gels; D006801:Humans; D007475:Ion Exchange Resins / Q000737:chemistry; D013058:Mass Spectrometry; D009705:Nucleosides / Q000302:isolation & purification* / Q000652:urine
D000180:Acrylic Resins; D003911:Dextrans; D005782:Gels; D007475:Ion Exchange Resins; D009705:Nucleosides; C016680:polyacrylamide gels; C025614:sephadex