Advertisement

High Performance Liquid Chromatography of Plasma Pyrimidines and Purines and Its Application in Cancer Chemotherapy

  • A. Leyva
  • J. Schornagel
  • H. M. Pinedo
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 122B)

Abstract

A variety of sensitive high performance liquid chromatography (HPLC) methods have been reported for the separation of pyrimidine and purine compounds1–4. These methods have involved anion- and cation-exchange and reverse phase chromatography. The use of these common techniques for the analysis of plasma is sometimes hampered by the incomplete fractionation of bases and nucleosides or inadequate separation of these compounds from other plasma components. Also, in most cases elution has required more than one buffer or the use of a gradient. Eksteen et al.5 recently demonstrated that bases and nucleosides could be rapidly and efficiently separated by HPLC using an anion-exchange resin and isocratic elution with an alcoholic phosphate buffer. Optimal results were reported achieved using 0.05 M sodium phosphate-0.005 M citric acid, pH 9.25, in 55% ethanol as the eluent and a column temperature of 70°. We initially attempted to use the same method for the analysis of plasma for pyrimidines and purines. However, it was noted that uric acid was not well separated from oxypurines. Moreover, using the conditions described above, we encountered poor column stability and inconsistent flow rates. By decreasing the phosphate and ethanol concentrations in the eluent, it was possible to obtain good column stability.

Keywords

High Performance Liquid Chromatography Uric Acid Isocratic Elution Reverse Phase Chromatography Phase High Performance Liquid Chromatography 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    R. P. Singhal and W. E. Cohn, Cation-Exchange Chromatography on Anion Exchanges: Application to Nucleic Acid Components and Comparison with Anion-Exchange Chromatography, Biochemistry 12:1532 (1973).PubMedCrossRefGoogle Scholar
  2. 2.
    P. R. Brown, S. Bobick and F. L. Hanley, The Analysis of Purine and Pyrimidine Bases and their Nucleosides by High-Pressure Liquid Chromatography, J. Chromatog. 99:587 (1974).CrossRefGoogle Scholar
  3. 3.
    F. S. Anderson and R. C. Murphy, Isocratic Separation of Some Purine Nucleotide, Nucleoside and Base Metabolites from Biological Extracts by High-Performance Liquid Chromatography, J. Chromatog. 121:251 (1976).CrossRefGoogle Scholar
  4. 4.
    A. M. Krstulovic, P. R. Brown and D. M. Rosle, Identification of Nucleosides and Bases in Serum and Plasma Samples by Reverse Phase High Performance Liquid Chromatography, Anal. Biochem. 49:2237 (1977).Google Scholar
  5. 5.
    R. Eksteen, J. C. Kraak and P. Linssen, Conditions for Rapid Separations of Nucleobases and Nucleosides by High-Pressure Anion-Exchange Chromatography, J. Chromatog. 148:413 (1978).CrossRefGoogle Scholar
  6. 6.
    W. D. Ensminger and E. Frei III, The Prevention of Methotrexate Toxicity by Thymidine Infusions in Humans, Cancer Res. 37: 1857 (1977).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • A. Leyva
    • 1
    • 2
  • J. Schornagel
    • 1
    • 2
  • H. M. Pinedo
    • 1
    • 2
  1. 1.Section of Experimental Chemotherapy, Antoni van Leeuwenhoek Institute and Department of OncologyFree University HospitalAmsterdam (A.L., H.M.P.)Netherlands
  2. 2.Department of Internal MedicineState University HospitalUtrecht (J.S.)The Netherlands

Personalised recommendations