Laboratory of Toxicology, Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium; Department of Laboratory Medicine, Ghent University Hospital, Ghent, Belgium.
Laboratory of Toxicology, Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
J Pharm Biomed Anal. 2022 Jan 5;207:114418. doi: 10.1016/j.jpba.2021.114418. Epub 2021 Oct 6.
Therapeutic drug monitoring (TDM) of tyrosine kinase inhibitors (TKIs) shows significant potential in guiding personalized anticancer treatment. Dried blood microsampling could be a valuable alternative for traditional plasma sampling to provide TDM results faster and to reach a wider audience. Sample collection is easy and patient friendly as only a small volume of blood is collected via a fingerprick. This enables the possibility of home sampling by the patients themselves. Therefore, an LC-MS/MS method was developed and validated for the quantification of bosutinib, dasatinib, gilteritinib, ibrutinib, imatinib, midostaurin, nilotinib and ponatinib in dried blood samples collected via volumetric absorptive microsampling (VAMS). A VAMS device collects a fixed volume of blood (± 10 µL), irrespective of the sample's hematocrit (Hct). During method validation, special attention was paid to the possible impact of Hct (range 0.18-0.55) on matrix effect (ME), robustness of the extraction, and accuracy of the method. The method was successfully validated based on international guidelines in terms of calibration curves, precision (within-run CV 2.20-14.8%; between-run CV 2.40-12.3%), accuracy (within-run bias 0.34-12.5%; between-run bias -0.15 to 16.2%), carry-over and selectivity. IS-compensated ME and recovery were Hct independent and no significant impact of Hct on the accuracy of the TKI quantifications was observed. All TKIs were stable in VAMS samples stored at -20 °C, 4 °C and room temperature for at least 4 weeks and for 2 days at 60 °C (except ibrutinib). Lastly, we demonstrated a good agreement between liquid blood obtained from patients on TKI treatment and VAMS samples prepared from that venous blood. As this implies that there is no methodological impact of liquid versus dried blood analysis, the presented method can be applied in clinical follow-up studies for determining TKIs in (capillary) VAMS samples with varying Hct levels.
治疗药物监测 (TDM) 的酪氨酸激酶抑制剂 (TKI) 在指导个体化抗癌治疗方面显示出巨大潜力。 干血微采样可以作为传统血浆采样的有价值替代方法，更快地提供 TDM 结果，并覆盖更广泛的受众。 样本采集简单且对患者友好，只需通过指尖采集少量血液。 这使得患者自己在家中采样成为可能。 因此，开发并验证了一种 LC-MS/MS 方法，用于定量分析通过体积吸收微采样（VAMS）采集的干血样中的博舒替尼、达沙替尼、吉特替尼、伊布替尼、伊马替尼、米哚妥林、尼洛替尼和泊那替尼。 VAMS 装置采集固定体积的血液（±10 μL），而与样本的血细胞比容（Hct）无关。 在方法验证过程中，特别关注 Hct（范围 0.18-0.55）对基质效应（ME）、提取的稳健性以及方法的准确性的可能影响。 根据国际指南，该方法在校准曲线、精密度（批内 CV 2.20-14.8%；批间 CV 2.40-12.3%）、准确度（批内偏差 0.34-12.5%；批间偏差-0.15 至 16.2%）、残留和选择性方面成功得到验证。 内标补偿 ME 和回收率与 Hct 无关，未观察到 Hct 对 TKI 定量准确性的显著影响。 所有 TKI 在 VAMS 样本中均稳定，在-20°C、4°C 和室温下至少可稳定 4 周，在 60°C 下可稳定 2 天（伊布替尼除外）。 最后，我们证明了在接受 TKI 治疗的患者的静脉血中获得的液态血与从该静脉血制备的 VAMS 样本之间存在良好的一致性。 这意味着液体血与干血分析之间没有方法学影响，因此，所提出的方法可应用于临床随访研究，以确定具有不同 Hct 水平的（毛细血管）VAMS 样本中的 TKI。
