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
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
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DOI：
10.1002/bmc.2741
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发表时间：
2013-01-01
影响因子：
1.8
通讯作者：
Largiader, Carlo R.
中科院分区：
文献类型：
--
作者：
Buechel, Barbara;Rhyn, Peter;Largiader, Carlo R.
化疗药物5-氟尿嘧啶（5-FU）被广泛用于治疗实体瘤。对5-FU治疗的反应各不相同，1030%的患者出现严重毒性，部分原因是二氢嘧啶脱氢酶（DPD）活性降低。DPD将内源性尿嘧啶(U)转化为5,6-二氢尿嘧啶（UH2），类似地，5- fu转化为5-氟-5,6-二氢尿嘧啶（5- fuh2）。U和UH2与5-FU和5-FUH2的联合定量可以提供DPD活性的治疗前评估，并进一步指导治疗期间的药物剂量。在这里，我们报道了一种液相色谱-串联质谱法同时定量人血浆中U， UH2, 5-FU和5-FUH2的发展。以10:1乙酸乙酯-2-丙醇（v/v）液液萃取制备样品。蒸发后的样品在0.1%甲酸中重构，10 μ L的等分液注入高效液相色谱系统。分析物在Atlantis dC18色谱柱上分离，流动相为1.0 mm乙酸铵、0.5 mm甲酸和3.3%甲醇。通过多重反应监测检测正离子分析物。在0.0110 μ m U、0.110 μ m UH2、0.175 μ m 5-FU和0.7575 μ m 5-FUH2的分析响应范围内呈线性，覆盖了血浆中预期的浓度范围。该方法按照FDA指南进行了验证，并应用于从10名5- fu治疗的结直肠癌患者中获得的临床样本。本方法将5-FU药代动力学和DPD活性的分析合并为一个单一的分析，代表了一个有价值的工具，以提高5-FU为基础的化疗的有效性和安全性。版权所有：John Wiley & Sons， Ltd。
The chemotherapeutic drug 5-fluorouracil (5-FU) is widely used for treating solid tumors. Response to 5-FU treatment is variable with 1030% of patients experiencing serious toxicity partly explained by reduced activity of dihydropyrimidine dehydrogenase (DPD). DPD converts endogenous uracil (U) into 5,6-dihydrouracil (UH2), and analogously, 5-FU into 5-fluoro-5,6-dihydrouracil (5-FUH2). Combined quantification of U and UH2 with 5-FU and 5-FUH2 may provide a pre-therapeutic assessment of DPD activity and further guide drug dosing during therapy. Here, we report the development of a liquid chromatographytandem mass spectrometry assay for simultaneous quantification of U, UH2, 5-FU and 5-FUH2 in human plasma. Samples were prepared by liquidliquid extraction with 10:1 ethyl acetate-2-propanol (v/v). The evaporated samples were reconstituted in 0.1% formic acid and 10 mu L aliquots were injected into the HPLC system. Analyte separation was achieved on an Atlantis dC18 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 0.0110 mu m for U, 0.110 mu m for UH2, 0.175 mu m for 5-FU and 0.7575 mu m for 5-FUH2, 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. Copyright (c) 2012 John Wiley & Sons, Ltd.