Determination of thymidine phosphorylase activity by a non-radiochemical assay using reversed-phase high-performance liquid chromatography.
Thymidine phosphorylase (TP) catalyses the conversion of thymidine into thymine. A non-radiochemical assay procedure for TP was developed in which thymine was detected at 265 nm after separation with reversed-phase HPLC. A complete separation of thymidine and thymine was achieved in 6 min and the minimum amount of thymine that could be detected was 0.8 pmol. The assay was linear with reaction times, up to at least 4 h, and protein concentrations up to at least 65 microg/ml. Population analysis showed no differences in TP activity between man and women or with increasing age.
van Kuilenburg
André B P
AB
Academic Medical Center, University of Amsterdam, Emma Children's Hospital and Department of Clinical Chemistry, 1100 DE Amsterdam, The Netherlands. a.b.vanKuilenburg@amc.uva.nl
Zoetekouw
Lida
L
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007700:Kinetics; D007962:Leukocytes / Q000201:enzymology*; D011854:Radiochemistry; D013939:Thymidine Phosphorylase / Q000378:metabolism*
D013939:Thymidine Phosphorylase