RESUMO
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%.
Assuntos
Nucleosídeos/isolamento & purificação , Resinas Acrílicas/química , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Dextranos/química , Géis , Humanos , Resinas de Troca Iônica/química , Espectrometria de Massas , Nucleosídeos/urinaRESUMO
Analysis of urine from cancer patients by capillary gas chromatography/mass spectrometry positively identified 14 urinary nucleosides including several modified nucleosides. Levels of the modified nucleosides 1-methyl-adenosine, 2-methylguanosine, N2,N2-dimethylguanosine and 1-methylinosine as well as the total nucleoside level were elevated in the urine when a malignant tumour was present; the levels of N2,N2-dimethylguanosine were found to correlate with the stage of the cancer.