A sensitive non-radioactive assay for pyrimidine nucleoside phosphorylase using reversed-phase high performance liquid chromatography

Emile J. Laurensse, Herbert M. Pinedo, Godefridus J. Peters

Research output: Contribution to journalArticlepeer-review

Abstract

A new sensitive assay for pyrimidine nucleoside phosphorylase using non-radioactive substrates is described. With the natural substrate undine (UR) and the analog, 5′-deoxy-5-fluorouridine (5′dFUR) conditions have been optimized to measure the product formation with reversed-phase high-performance liquid chromatography. Using automated injection large series of samples may be analyzed. The assay for UR phosphorylase appeared to be comparable to existing methods with radiolabeled UR as substrate regarding sensitivity and linearity. The assay has been used to measure kinetic parameters for 5′dFUR and UR in two cell lines from intestinal origin.

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalClinica Chimica Acta
Volume178
Issue number1
DOIs
StatePublished - 1988
Externally publishedYes

Keywords

  • 5′-Deoxy-5-fluorouridine
  • High-performance liquid chromatography
  • Pyrimidine nucleoside phosphorylase
  • Uridine

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

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