Department of Clinical Pharmacy, Oita University Hospital, 1-1 Idaigaoka, Hasama-machi, Yufu-shi, Oita, 879-5593, Japan.
Department of Gastroenterological and Pediatric Surgery, Faculty of Medicine, Oita University, Yufu-shi, Oita, Japan.
Sci Rep. 2021 Feb 4;11(1):3132. doi: 10.1038/s41598-021-82908-8.
Combination therapy of tegafur/uracil (UFT) and leucovorin (LV) is widely used to treat colorectal cancers. Although this therapy has a significant therapeutic effect, severe adverse effects occur frequently. Therapeutic drug monitoring (TDM) may help to prevent adverse effects. A useful assay that can quantitate plasma levels of 5-FU, uracil, and tegafur simultaneously for TDM has been desired, but such a method is not currently available. In this study, we aimed to develop a sensitive method for simultaneous quantification of 5-FU, uracil, and tegafur in human plasma using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). After preparing plasma samples by protein precipitation and liquid extraction, 5-FU, uracil, and tegafur were analyzed by UPLC-MS/MS in negative electrospray ionization mode. Validation was performed according to US Food and Drugs Administration guidance. The calibration curves were linear over concentration ranges of 2-500 ng/mL for 5-FU, 20-5000 ng/mL for uracil, and 200-50,000 ng/mL for tegafur. The corresponding average recovery rates were 79.9, 80.9, and 87.8%. The method provides accuracy within 11.6% and precision below 13.3% for all three analytes. Matrix effects of 5-FU, uracil, and tegafur were higher than 43.5, 84.9, and 100.2%, respectively. This assay was successfully applied to assess the time courses of plasma 5-FU, uracil, and tegafur concentrations in two patients with colorectal liver metastasis who received UFT/LV therapy after hepatectomy. In conclusion, we succeeded to develop a sensitive and robust UPLC-MS/MS method for simultaneous quantification of 5-FU, uracil, and tegafur in human plasma. This method is potentially useful for TDM in patients receiving UFT/LV combination therapy.
替加氟/尿嘧啶（UFT）联合亚叶酸（LV）的联合治疗被广泛用于治疗结直肠癌。尽管这种治疗方法具有显著的治疗效果，但经常会出现严重的不良反应。治疗药物监测（TDM）可能有助于预防不良反应。人们一直希望有一种有用的测定方法，可以同时定量测定血浆中的 5-FU、尿嘧啶和替加氟，用于 TDM，但目前还没有这样的方法。在这项研究中，我们旨在开发一种灵敏的方法，用于同时定量测定人血浆中的 5-FU、尿嘧啶和替加氟，采用超高效液相色谱-串联质谱法（UPLC-MS/MS）。通过蛋白沉淀和液液萃取制备血浆样品后，采用负电喷雾电离模式的 UPLC-MS/MS 分析 5-FU、尿嘧啶和替加氟。验证符合美国食品和药物管理局的指导原则。5-FU、尿嘧啶和替加氟的校准曲线在 2-500ng/mL 浓度范围内呈线性，5-FU、尿嘧啶和替加氟的平均回收率分别为 79.9%、80.9%和 87.8%。该方法对所有三种分析物的准确度均在 11.6%以内，精密度均在 13.3%以下。5-FU、尿嘧啶和替加氟的基质效应分别高于 43.5%、84.9%和 100.2%。该方法成功应用于评估两例结直肠癌肝转移患者行肝切除术后接受 UFT/LV 治疗的血浆 5-FU、尿嘧啶和替加氟浓度的时间过程。总之，我们成功开发了一种灵敏而强大的 UPLC-MS/MS 方法，用于同时定量测定人血浆中的 5-FU、尿嘧啶和替加氟。该方法对于接受 UFT/LV 联合治疗的患者的 TDM 具有潜在的应用价值。
