Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells

Dick Pluim; Bart A. Jacobs; Maarten J. Deenen; Anneloes E. Ruijter; Robin M. Van Geel; Artur M. Burylo; Didier Meulendijks; Jos H. Beijnen; Jan H. Schellens

doi:10.4155/bio.14.304

Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells

Dick Pluim, Bart A. Jacobs, Maarten J. Deenen, Anneloes E. Ruijter, Robin M. Van Geel, Artur M. Burylo, Didier Meulendijks, Jos H. Beijnen, Jan H. Schellens^*

^*Corresponding author for this work

Pharmacoepidemiology and Clinical Pharmacology

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Pages (from-to)	519-529
Number of pages	11
Journal	Bioanalysis
Volume	7
Issue number	5
DOIs	https://doi.org/10.4155/bio.14.304
Publication status	Published - 1 Mar 2015

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title = "Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells",

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.",

author = "Dick Pluim and Jacobs, {Bart A.} and Deenen, {Maarten J.} and Ruijter, {Anneloes E.} and {Van Geel}, {Robin M.} and Burylo, {Artur M.} and Didier Meulendijks and Beijnen, {Jos H.} and Schellens, {Jan H.}",

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doi = "10.4155/bio.14.304",

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journal = "Bioanalysis",

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T1 - Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells

AU - Pluim, Dick

AU - Jacobs, Bart A.

AU - Deenen, Maarten J.

AU - Ruijter, Anneloes E.

AU - Van Geel, Robin M.

AU - Burylo, Artur M.

AU - Meulendijks, Didier

AU - Beijnen, Jos H.

AU - Schellens, Jan H.

PY - 2015/3/1

Y1 - 2015/3/1

N2 - 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.

AB - 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.

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JO - Bioanalysis

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Improved pharmacodynamic assay for dihydropyrimidine dehydrogenase activity in peripheral blood mononuclear cells

Abstract

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