A selective, sensitive and rapid LC–MS/MS method has been developed and validated as per US Food and Drug Administration regulatory guidelines for the simultaneous quantitation of colchicine and febuxostat in rat plasma. Colchicine and febuxostat were extracted from the rat plasma using 10% tert‐butyl methyl ether in ethyl acetate using colchicine‐d₆ as an internal standard (IS). The chromatographic separation of colchicine, febuxostat and the IS was achieved using a mobile phase comprising 5 mm ammonium formate and 0.025% formic acid in acetonitrile (20:80, v/v) in isocratic mode on an Eclipse XDB‐C₁₈ column. The injection volume and flow rate were 5.0 μl and 0.9 ml/min, respectively. Colchicine and febuxostat were detected by positive electrospray ionization in multiple reaction monitoring mode using transition pairs (Q1 → Q3) of m/z 400.10 → 358.10 and 317.05 → 261.00, respectively. The assay was linear in the ranges of 0.25–254 and 2.60–622 ng/ml for colchicine and febuxostat, respectively. The inter‐ and intra‐day precision values were 0.58–13.0 and 1.03–4.88% for colchicine and febuxostat, respectively. No matrix or carryover effects were observed during the validation. Both analytes were stable on the bench‐top, in the autosampler and in storage (freeze–thaw cycles and long‐term storage at −80°C). A pharmacokinetic study in rats was performed to show the applicability of the validated method.

中文翻译：

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在大鼠血浆中同时测定秋水仙碱和非布索坦：在大鼠药代动力学研究中的应用。

一种选择性，灵敏和快速的LC-MS / MS方法已经根据美国食品和药物管理局的监管指南进行了开发和验证，可同时定量大鼠血浆中的秋水仙碱和非布索坦。使用秋水仙碱d ₆作为内标（IS），使用10％的叔丁基甲基醚的乙酸乙酯溶液从大鼠血浆中提取秋水仙碱和非布索坦。秋水仙碱，非布索坦和IS的色谱分离是通过在Eclipse XDB-C ₁₈上等度模式下使用包含5 m m甲酸铵和0.025％甲酸的乙腈溶液（20:80，v / v）进行的。柱。进样量和流速分别为5.0μl和0.9 ml / min。在多个反应监测模式下，通过正电喷雾电离分别使用m / z 400.10→358.10和317.05→261.00的过渡对（Q1→Q3）检测秋水仙碱和非布索坦。秋水仙碱和非布索坦的测定线性范围分别为0.25–254和2.60–622 ng / ml。秋水仙碱和非布索坦的日间和日内精度值分别为0.58–13.0和1.03–4.88％。验证期间未观察到基质或残留效应。两种分析物在台式，自动进样器和储存中均稳定（冻融循环和在-80 ° C下长期储存）C）。在大鼠中进行了药代动力学研究，以证明该方法的适用性。