Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells
Publication date
2015-03-01
Authors
Pluim, Dick
Jacobs, Bart A.
Deenen, Maarten J.
Ruijter, Anneloes E.
Van Geel, Robin M.
Burylo, Artur M.
Meulendijks, Didier
Beijnen, Jos H.
Schellens, Jan H M
Editors
Advisors
Supervisors
Document Type
Article
Metadata
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License
taverne
Abstract
Background: Dihydropyrimidine dehydrogenase (DPD) activity determination in peripheral blood mononuclear cells of DPD deficient patients was hitherto inaccurate due to hemoglobin (Hb) contamination. We developed an improved method for accurate measurement of DPD activity in patients. Results: DPD activity was determined by HPLC with online radioisotope detection using liquid scintillation counting. Hb was determined spectrophotometrically. Method accuracy and precision were significantly improved by using cumulative area of all peaks as IS. Peripheral blood mononuclear cell lysates from DPD deficient patients were highly contaminated with on average 23.3% (range 2.7-51%) of Hb resulting in up to twofold underestimated DPD activity. DPD activities were corrected for Hb contamination. The method was validated and showed good long-term sample stability. Conclusion: This method has increased specificity allowing accurate identification of DPD deficient patients.
Keywords
Taverne, Clinical Biochemistry, Pharmacology, Toxicology and Pharmaceutics(all), Medical Laboratory Technology, Analytical Chemistry, General Medicine
Citation
Pluim, D, Jacobs, B A, Deenen, M J, Ruijter, A E, Van Geel, R M, Burylo, A M, Meulendijks, D, Beijnen, J H & Schellens, J H 2015, 'Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells', Bioanalysis, vol. 7, no. 5, pp. 519-529. https://doi.org/10.4155/bio.14.304