Ibrutinib is an orally administered first-in-class irreversible Bruton's tyrosine kinase (BTK) covalent inhibitor for the treatment of patients with B-cell malignancies. Several isolated clinical observations reported its efficacy in central nervous system dissemination. Herein, we described the development and validation of an ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) procedure for the quantification of ibrutinib and its active metabolite PCI-45227 in cerebrospinal fluid (CSF). This is the first complete validated method for quantification of ibrutinib and PCI-45227 in CSF. The compounds were eluted on a Waters BEH C18 column (50.0 × 2.1 mm; 1.7 μm) using a gradient elution with a mobile phase composed of ammonium formate buffer 5 mM pH 3.2 and acetonitrile +0.1% formic acid with a flow rate of 400 μL·min⁻¹. Two deuterated internal standards were used to obtain the most accurate quantification. The CSF samples were prepared by a simple and rapid dilution. The method was validated by testing the selectivity, response function, intra-day and inter-day precisions, trueness, limits of detection (LOD) and lower limits of quantification (LLOQ). The validation results proved that the methods were suitable to quantify ibrutinib and PCI-45227 in real biological CSF samples from 0.50 (ibrutinib) or 1.00 (PCI-45227) to 30.00 ng·mL⁻¹. Furthermore, the developed method was adapted to allow the quantification of both compounds in plasma and the results were compared to those reported in literature. The plasmatic samples were treated by protein precipitation and the method was validated to quantify ibrutinib and PCI-45227 in real biological plasmatic samples from 5.00 to 491 ng·mL⁻¹. Lastly, for both matrices, accuracy profiles were plotted from the trueness and precision results using a 20% α-risk (β = 80%) and the tolerance intervals were comprised within the acceptance limits fixed at ±25% for the LLOQ and ±15% for the other concentrations. Finally, these methods were successfully applied to quantify ibrutinib and PCI-45227 in real human CSF and plasma samples.

依鲁替尼是一种口服的不可逆的布鲁顿酪氨酸激酶（BTK）共价抑制剂，用于治疗B细胞恶性肿瘤。几项孤立的临床观察报告了其在中枢神经系统传播中的功效。在此，我们描述了用于定量分析脑脊液（CSF）中依鲁替尼及其活性代谢物PCI-45227的超高效液相色谱-串联质谱（UHPLC-MS / MS）程序的开发和验证。这是第一个用于脑脊液中依鲁替尼和PCI-45227定量的完整验证方法。将化合物在Waters BEH C18色谱柱（50.0×2.1 mm; 1.7μm）上进行洗脱，并使用流动相进行洗脱，该流动相由甲酸铵缓冲液5 mM pH 3.2和乙腈+ 0.1％甲酸组成，流速为400μL ·min^-1。使用两个氘代内标获得最准确的定量。通过简单而快速的稀释来制备CSF样品。通过测试选择性，响应函数，日内和日间精度，真实性，检测限（LOD）和定量下限（LLOQ）验证了该方法。验证结果证明，该方法适合定量从0.50（依鲁替尼）或1.00（PCI-45227）到30.00 ng·mL ^-1的真实生物CSF样品中的依鲁替尼和PCI-45227。此外，已开发的方法适用于定量血浆中的两种化合物，并将结果与​​文献报道的结果进行了比较。对血浆样品进行蛋白质沉淀处理，并验证了该方法的定量结果，该方法可定量测定从5.00到491 ng·mL ^-1的真实生物血浆样品中的依鲁替尼和PCI-45227 。最后，对于两个矩阵，使用20％的α风险（β= 80％）从真实性和精确度结果中绘制出精度分布图，并且容差区间包含在LLOQ和±15固定为±25％的接受范围内其他浓度的％。最后，这些方法已成功应用于定量真实人CSF和血浆样品中的依鲁替尼和PCI-45227。