Development of a purification procedure for the isolation of nucleosides from urine prior to mass spectrometric analysis |
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Authors: | Dudley E El-Shakawi S Games D E Newton R P |
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Institution: | Biochemistry Research Group, School of Biological Sciences, University of Wales Swansea, UK. |
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Abstract: | 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%. |
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