Quantitative high-performance liquid chromatography of nucleosides in biological materials

Charles W. Gehrke, Kenneth C. Kuo, George E. Davis, Robert D. Suits, T. Phillip Waalkes, Ernest Borek

Research output: Contribution to journalArticle

Abstract

A rigorous, comprehensive, and reliable reversed-phase high-performance liqiud chromatographic (HPLC) method has been developed for the analysis of ribonucleosides in urine (Ψ, m1A, m1I, m2G, A, m22G. An initial isolation of ribonucleosides with an affinity gel containing an immobilized phenylboronic acid was used to improve selectivity and sensitivity. Response for all nucleosides was linear from 0.1 to 50 nmoles injected and good quantitation was obtained for 25 μl or less of sample placed on the HPLC column. Excellent precision of analysis for urinary nucleosides was achieved on matrix dependent and independent samples, and the high resolution of the reversed-phase column allowed the complete separation of 9 nucleosides from other unidentified UV absorbing components at the 1-ng level. Supporting experimental data are presented on precision, recovery, chromatographic methods, minimum detection limit, retention time, relative molar response, sample clean-up, stability of nucleosides, boronate gel capacity, and application to analysis of urine from patients with leukemia and breast cancer. This method is now being used routinely for the determination of the concentration and ratios of nucleosides in urine from patients with different types of cancer and in chemotherapy response studies.

Original languageEnglish (US)
Pages (from-to)455-476
Number of pages22
JournalJournal of Chromatography A
Volume150
Issue number2
DOIs
StatePublished - Mar 21 1978

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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  • Cite this

    Gehrke, C. W., Kuo, K. C., Davis, G. E., Suits, R. D., Waalkes, T. P., & Borek, E. (1978). Quantitative high-performance liquid chromatography of nucleosides in biological materials. Journal of Chromatography A, 150(2), 455-476. https://doi.org/10.1016/S0021-9673(00)88205-9