A UHPLC-MS/MS method for the simultaneous quantification of BTK inhibitors and their active metabolite in human plasma and cerebrospinal fluid.

IF 2.1 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2026-07-01 Epub Date: 2026-07-15 DOI:10.1080/17576180.2026.2700356

Sijia Li, Jiameng Xie, Fan Xia, Tingting Yang, Fangyue Zheng, Liyan Miao, Chenrong Huang

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引用次数: 0

Abstract

Background: Bruton's tyrosine kinase inhibitors (BTKis, ibrutinib, zanubrutinib, orelabrutinib) are key targeted therapies for B-cell lymphomas, but interindividual variability and adverse reactions limit their use. Metabolized by CYP3A, co-medications or hepatic dysfunction cause plasma fluctuations, necessitating therapeutic drug monitoring (TDM). Cerebrospinal fluid (CSF) distribution is also critical for CNS lymphoma treatment.

Objective: To develop and validate a UHPLC-MS/MS method for simultaneous quantification of three BTKis and one metabolite in human plasma and CSF to support TDM.

Methods: After protein precipitation with methanol, samples were analyzed on an ACQUITY HSS T3 column with a 3-min gradient at 0.3 mL/min and ESI (+) MRM. Mobile phase: water-acetonitrile-formic acid-ammonium acetate (1 M) (950:50:1:1, A) and (50:950:1:1, B).

Results: The method showed excellent selectivity and linearity (r > 0.997) over 1-200 ng/mL (ibrutinib, zanubrutinib, ibrutinib metabolite) and 5-1000 ng/mL (orelabrutinib) in both matrices. Recoveries were 89.4-103.8%, matrix factors 0.93-1.06, and precision CV ≤8.5%. Stability was confirmed. The method was applied to 34 clinical samples (25 plasma, 9 CSF) from 20 patients.

Conclusion: This method detects four BTK-related analytes in plasma/CSF with a 3-min run time, evaluation of hemolytic/lipemic matrices, and clinical validation, increasing throughput, reducing costs, and broadening applicability, providing preliminary feasibility data for clinical TDM.

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UHPLC-MS/MS同时定量人血浆和脑脊液中BTK抑制剂及其活性代谢物的方法

背景：布鲁顿酪氨酸激酶抑制剂（BTKis、依鲁替尼、zanubrutinib、orelabrutinib）是b细胞淋巴瘤的关键靶向治疗药物，但个体间差异和不良反应限制了它们的使用。由CYP3A代谢，联合用药或肝功能障碍引起血浆波动，需要治疗性药物监测（TDM）。脑脊液（CSF）的分布对于中枢神经系统淋巴瘤的治疗也是至关重要的。目的：建立并验证同时定量人血浆和脑脊液中3种BTKis和1种代谢物的UHPLC-MS/MS方法，以支持TDM。方法：用甲醇沉淀蛋白质后，样品在ACQUITY HSS T3柱上进行分析，梯度为0.3 mL/min, ESI (+) MRM。流动相：水-乙腈-甲酸-乙酸铵（1 M）（950:50:1:1， A）和（50:950:1:1，B）。结果：该方法在1 ~ 200 ng/mL（依鲁替尼、扎鲁替尼、依鲁替尼代谢物）和5 ~ 1000 ng/mL（奥瑞鲁替尼）范围内均具有良好的选择性和线性（r为0.9997）。加样回收率为89.4 ~ 103.8%，基质因子0.93 ~ 1.06，精密度CV≤8.5%。稳定性得到证实。该方法应用于20例患者的34份临床样本（25份血浆，9份脑脊液）。结论：该方法检测血浆/脑脊液中4种btk相关分析物，运行时间为3 min，对溶血/血脂基质进行评估，并进行临床验证，提高了吞吐量，降低了成本，扩大了适用性，为临床TDM提供了初步的可行性数据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.