PMID: 7655619Apr 7, 1995Paper

Purine enzyme activities in peripheral blood mononuclear cells: comparison of a new non-radiochemical high-performance liquid chromatography procedure and a radiochemical thin-layer chromatography procedure

Journal of Chromatography. B, Biomedical Applications
J N StolkL B van de Putte

Abstract

Purine enzyme activities are usually assayed by radiochemical procedures and often TLC is part of the separation method. In screening patients with rheumatic diseases, these procedures have shown disadvantages like a relatively large coefficient of variation (C.V.) and time-instability. We describe a non-radiochemical reversed-phase HPLC micro-method with UV detection for measurement of activities of purine 5'-nucleotidase (5'NT; EC 3.1.3.5), purine nucleoside phosphorylase (PNP; EC 2.4.2.1) and hypoxanthine guanine phosphoribosyltransferase (HGPRT; EC 2.4.2.8) in human peripheral blood mononuclear cells (PBMC). The HPLC procedure is compared with the radiochemical TLC procedure by testing both with a 5'NT and a PNP assay. Reproducibility is tested with 14 healthy controls in each procedure. Short-term and long-term time-stability is tested by comparing enzyme activities measured immediately after preparation of the PBMC (week 0) with those found after freezing and storage at -20 degrees C for a maximum of 10 weeks. The HPLC procedure is preferable to the radiochemical TLC procedure because it shows significantly better reproducibility and better time-stability and in addition is non-radiochemical and less time-consuming.

Citations

May 30, 2009·European Journal of Applied Physiology·Jacek ZielińskiMaria Laurentowska
Sep 12, 2012·European Journal of Applied Physiology·Jacek ZielińskiEwa Słomińska
Jun 9, 1995·Biochimica Et Biophysica Acta·M TavenierJ W De Jong
Mar 7, 2002·Clinica Chimica Acta; International Journal of Clinical Chemistry·Heide DaxeckerMathias M Müller
Nov 14, 2002·Annals of the Rheumatic Diseases·A E van EdeL B van de Putte
Oct 23, 2013·Analytical Biochemistry·Marianne FreundliebChrista E Müller
Dec 20, 2005·The Kaohsiung Journal of Medical Sciences·Shun-Jen ChangYing-Chin Ko
Jun 19, 2014·Nucleosides, Nucleotides & Nucleic Acids·Anna WojtuszkiewiczG J Peters
Dec 14, 2011·Journal of Applied Physiology·Jacek Zieliński, Krzysztof Kusy
Mar 3, 2011·Medicine and Science in Sports and Exercise·Jacek ZielińskiTadeusz Rychlewski
Mar 29, 1996·Journal of Chromatography. B, Biomedical Applications·C W Keuzenkamp-JansenJ M Trijbels
Jun 5, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·H WardJ E Foker

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