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

Fingerprint

High performance liquid chromatography
Nucleosides
Biological materials
High Pressure Liquid Chromatography
Ribonucleosides
Urine
Gels
Chemotherapy
Limit of Detection
Leukemia
Breast Neoplasms
Drug Therapy
Recovery
Neoplasms

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry
  • Molecular Medicine

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

Quantitative high-performance liquid chromatography of nucleosides in biological materials. / Gehrke, Charles W.; Kuo, Kenneth C.; Davis, George E.; Suits, Robert D.; Waalkes, T. Phillip; Borek, Ernest.

In: Journal of Chromatography A, Vol. 150, No. 2, 21.03.1978, p. 455-476.

Research output: Contribution to journalArticle

Gehrke, CW, Kuo, KC, Davis, GE, Suits, RD, Waalkes, TP & Borek, E 1978, 'Quantitative high-performance liquid chromatography of nucleosides in biological materials', Journal of Chromatography A, vol. 150, no. 2, pp. 455-476. https://doi.org/10.1016/S0021-9673(00)88205-9
Gehrke, Charles W. ; Kuo, Kenneth C. ; Davis, George E. ; Suits, Robert D. ; Waalkes, T. Phillip ; Borek, Ernest. / Quantitative high-performance liquid chromatography of nucleosides in biological materials. In: Journal of Chromatography A. 1978 ; Vol. 150, No. 2. pp. 455-476.
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