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

被引：0
|
作者：
Pluim, Dick [1 ]
Jacobs, Bart A. W. [1 ]
Deenen, Maarten J. [1 ]
Ruijter, Anneloes E. M. [1 ]
van Geel, Robin M. J. M. [1 ]
Burylo, Artur M. [1 ]
Meulendijks, Didier [1 ]
Beijnen, Jos H. [2 ,3 ]
Schellens, Jan H. M. [1 ,3 ]
机构：
[1] Netherlands Canc Inst, Div Mol Pathol, NL-1066 CX Amsterdam, Netherlands
[2] Netherlands Canc Inst, Dept Pharm & Pharmacol, NL-1066 EC Amsterdam, Netherlands
[3] Univ Utrecht, Fac Sci, Dept Pharmaceut Sci, Div Pharmacoepidemiol & Clin Pharmacol, NL-3584 CA Utrecht, Netherlands
来源：
BIOANALYSIS | 2015年 / 7卷 / 05期
关键词：
TOXICITY; CANCER; FLUOROURACIL; DEFICIENCY; DPYD; CAPECITABINE; EFFICACY; GENE;
D O I：
10.4155/BIO.14.304
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：519 / 529
页数：11
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