High-performance liquid chromatographic assay with ultraviolet detection for quantification of dihydrofluorouracil in human lymphocytes: application to measurement of dihydropyrimidine dehydrogenase activity

Deporte-Fety, R.; Picot, Marie Carene Nancy; Amiand, Marie-Bernadette; Moreau, A.; Campion, Loı̈c; Lanoë, Didier; Renée, Nicole; Milano, Gérard

doi:10.1016/s0378-4347(01)00359-0

Cited by 21 publications

(15 citation statements)

References 12 publications

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“…According to the recently published results from Remaud et al [18], plasmas were immediately separated by centrifugation at +4 • C and stored at −70 • C, in order to avoid changes in U concentration. PBMC DPD activity was measured according to the method of Johnson et al [8] modified by Deporte et al [9]. PBMC cells were isolated from 15 mL heparinized anticoagulant blood.…”

Section: Patients Plasma Analysismentioning

confidence: 99%

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High-performance liquid chromatographic assay with UV detection for measurement of dihydrouracil/uracil ratio in plasma

Déporte¹,

Amiand²,

Moreau³

et al. 2006

Journal of Chromatography B

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Section: Patients Plasma Analysismentioning

confidence: 99%

“…Direct methods are based on the determination of the precise DPD enzymatic activity in PBMC [8,9]. These methods require PBMC isolation and high-performance liquid chromatography (HPLC) analysis of 5,6-dihydrofluorouracil (5-FUH2) formed from 5-FU, making the procedure timeconsuming and labor-intensive.…”

Section: Introductionmentioning

confidence: 99%

High-performance liquid chromatographic assay with UV detection for measurement of dihydrouracil/uracil ratio in plasma

Déporte¹,

Amiand²,

Moreau³

et al. 2006

Journal of Chromatography B

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“…As a result, the HPLC-MS/MS based assay allows for a short run time and high precision and accuracy. The analysis time of the HPLC-MS/MS assay is considerably shorter than that of HPLC-ultraviolet procedures and those using radiolabeled substrates (4,6,7 ). In addition, the detection limit of the HPLC-MS/MS assay is 5 times lower than that of the reversed-phase HPLC-ultraviolet procedure, in which the product 5-fluorodihydrouracil is detected at 205 nm (4 ).…”

confidence: 99%

HPLC-Electrospray Tandem Mass Spectrometry for Rapid Determination of Dihydropyrimidine Dehydrogenase Activity

Kuilenburg¹,

Lenthe

Zoetekouw

et al. 2007

Clinical Chemistry

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Background: Patients with a partial dihydropyrimidine dehydrogenase (DPD) deficiency have an increased risk of developing severe 5-fluorouracil–associated toxicity. We developed a rapid and specific method to measure the DPD activity in peripheral blood mononuclear cells using HPLC tandem-mass spectrometry (HPLC-MS/MS). Methods: The activity of DPD was measured with thymine as the substrate, followed by reversed-phase HPLC combined with electrospray ionization MS/MS and detection of the product dihydrothymine with multiple-reaction monitoring. Stable-isotope labeled dihydrothymine was used as the internal standard. Results: Dihydrothymine was measured within an analytical run of 10 min, with a lower limit of quantification of 54 μg/L (0.4 μmol/L). The intraassay and interassay variations of the DPD activity assay were both <7%. A linear correlation (R2 = 0.980; P <0.001) was observed between the HPLC-MS/MS data and those obtained with a reference method using radiolabeled thymine. There were no systematic differences between the 2 methods, and both methods yielded similar results. Conclusion: The analysis of the DPD activity with HPLC-MS/MS is rapid, accurate, and sufficiently sensitive to be used as a screening method for patients with a DPD deficiency.

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“…3) une approche biochimique permettant la mesure de l'activité enzymatique de la DPD leucocytaire a été développée [4,10,20] ; 4) une approche pharmacologique par dosage de l'uracile et/ou de la thymine plasmatique ou urinaire [1] ; 5) une approche pharmacologique par dosage plasmatique simultané de l'uracile et du dihydrouracile endogènes par Chromatographie Liquide à Haute Performance (CLHP) ou par Chromatographie Liquide couplée à la Spectrométrie de Masse en tandem (LC/MS-MS) afin d'établir le rapport UH 2 /U, reflet de l'activité globale de la DPD [13,14] ; 6) une adaptation individuelle de la dose de 5-FU par suivi pharmacocinétique [13,28].…”

Section: Le Gène De La Dpdunclassified

Pharmacogenetics of the metabolism of pyrimidines. Dihydropyrimidin dehydrogenase

2005

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High-performance liquid chromatographic assay with ultraviolet detection for quantification of dihydrofluorouracil in human lymphocytes: application to measurement of dihydropyrimidine dehydrogenase activity

Cited by 21 publications

References 12 publications

High-performance liquid chromatographic assay with UV detection for measurement of dihydrouracil/uracil ratio in plasma

High-performance liquid chromatographic assay with UV detection for measurement of dihydrouracil/uracil ratio in plasma

HPLC-Electrospray Tandem Mass Spectrometry for Rapid Determination of Dihydropyrimidine Dehydrogenase Activity

Pharmacogenetics of the metabolism of pyrimidines. Dihydropyrimidin dehydrogenase

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