Therapeutic drug monitoring is important in patients taking BCR-ABL and Bruton’s tyrosine kinase inhibitors (TKIs). Some TKI active metabolites with long elimination half-lives, such as dihydrodiol ibrutinib (DHI), N-desmethyl imatinib (N-DI), and N-desmethyl ponatinib (N-DP), have been characterized, indicating that these active metabolites should be monitored along with the parent compounds. However, there are currently no methods for the simultaneous quantification of BCR-ABL and Bruton’s TKIs and their three active metabolites. The present study aimed to develop and validate a method for the simultaneous quantification of nine pharmacologically active compounds (bosutinib, dasatinib, DHI, ibrutinib, imatinib, N-DI, N-DP, nilotinib, and ponatinib) using high-performance liquid chromatography–tandem mass spectrometry. A 150-μL sample of plasma was analyzed after purification with supported liquid extraction. The method has a run time of 7 min and was successfully validated over the following calibration ranges: 0.25-75 ng/mL for N-DP, 0.5-150 ng/mL for dasatinib and ponatinib, 10-3000 ng/mL for imatinib and nilotinib, and 1-300 ng/mL for the other analytes. Stability of the analytes after short- and long-term storage in the presence of plasma matrix was examined, and all analytes were found to be stable under all tested conditions. The recovery was ≥83%, and the relative standard deviation of internal-standard normalized matrix effects ranged from 3.9 to 13.9%. Dilution integrity up to 4-fold was ensured. The applicability of the method for all analytes was demonstrated using patient samples.

对于服用BCR-ABL和Bruton酪氨酸激酶抑制剂（TKIs）的患者，治疗药物监测非常重要。一些具有长消除半衰期的TKI活性代谢物，例如二氢二醇依鲁替尼（DHI），N-去甲基伊马替尼（N-DI）和Nβ-去甲基ponatinib（N-DP）已被表征，表明这些活性代谢物应与母体化合物一起进行监测。但是，目前尚无同时定量BCR-ABL和Bruton's TKI及其三种活性代谢物的方法。本研究旨在开发和验证一种使用高效液相色谱法同时定量分析九种药理活性化合物（波舒替尼，达沙替尼，DHI，依鲁替尼，伊马替尼，N-DI，N-DP，尼洛替尼和ponatinib）的方法–串联质谱 在支持的液体萃取纯化后，分析了150μL血浆样品。该方法的运行时间为7分钟，并在以下校准范围内成功验证：N-DP为0.25-75 ng / mL，达沙替尼和ponatinib为0.5-150 ng / mL，伊马替尼和尼洛替尼的浓度为10-3000 ng / mL，其他分析物的浓度为1-300 ng / mL。检查了在血浆基质存在下短期和长期保存后分析物的稳定性，发现所有分析物在所有测试条件下均稳定。回收率≥83％，内标标准化基质效应的相对标准偏差为3.9-13.9％。确保稀释完整性高达4倍。使用患者样品证明了该方法对所有分析物的适用性。内标归一化矩阵效应的相对标准偏差在3.9％至13.9％之间。确保稀释完整性高达4倍。使用患者样品证明了该方法对所有分析物的适用性。内标归一化矩阵效应的相对标准偏差在3.9％至13.9％之间。确保稀释完整性高达4倍。使用患者样品证明了该方法对所有分析物的适用性。