LC-MS/MS method for simultaneous analysis of uracil, 5,6-dihydrouracil, 5-fluorouracil and 5-fluoro-5,6-dihydrouracil in human plasma for therapeutic drug monitoring and toxicity prediction in cancer patients
| Autor: | Stefan Schürch, Claudia Bühr, Carlo R. Largiadèr, Barbara Büchel, Ursula Amstutz, Peter Rhyn |
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| Rok vydání: | 2012 |
| Předmět: |
Formic acid
Clinical Biochemistry 01 natural sciences Biochemistry High-performance liquid chromatography Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Pharmacokinetics Drug Discovery medicine Dihydropyrimidine dehydrogenase Molecular Biology Pharmacology Chromatography medicine.diagnostic_test 010401 analytical chemistry Selected reaction monitoring Dihydrouracil General Medicine 3. Good health 0104 chemical sciences chemistry Fluorouracil Therapeutic drug monitoring 030220 oncology & carcinogenesis medicine.drug |
| Zdroj: | Biomedical Chromatography. 27:7-16 |
| ISSN: | 0269-3879 |
| DOI: | 10.1002/bmc.2741 |
| Popis: | The chemotherapeutic drug 5-fluorouracil (5-FU) is widely used for treating solid tumors. Response to 5-FU treatment is variable with 10-30% of patients experiencing serious toxicity partly explained by reduced activity of dihydropyrimidine dehydrogenase (DPD). DPD converts endogenous uracil (U) into 5,6-dihydrouracil (UH(2) ), and analogously, 5-FU into 5-fluoro-5,6-dihydrouracil (5-FUH(2) ). Combined quantification of U and UH(2) with 5-FU and 5-FUH(2) may provide a pre-therapeutic assessment of DPD activity and further guide drug dosing during therapy. Here, we report the development of a liquid chromatography-tandem mass spectrometry assay for simultaneous quantification of U, UH(2) , 5-FU and 5-FUH(2) in human plasma. Samples were prepared by liquid-liquid extraction with 10:1 ethyl acetate-2-propanol (v/v). The evaporated samples were reconstituted in 0.1% formic acid and 10 μL aliquots were injected into the HPLC system. Analyte separation was achieved on an Atlantis dC(18) column with a mobile phase consisting of 1.0 mm ammonium acetate, 0.5 mm formic acid and 3.3% methanol. Positively ionized analytes were detected by multiple reaction monitoring. The analytical response was linear in the range 0.01-10 μm for U, 0.1-10 μm for UH(2) , 0.1-75 μm for 5-FU and 0.75-75 μm for 5-FUH(2) , covering the expected concentration ranges in plasma. The method was validated following the FDA guidelines and applied to clinical samples obtained from ten 5-FU-treated colorectal cancer patients. The present method merges the analysis of 5-FU pharmacokinetics and DPD activity into a single assay representing a valuable tool to improve the efficacy and safety of 5-FU-based chemotherapy. |
| Databáze: | OpenAIRE |
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