目的 采用超高效液相色谱-串联质谱（ UPLC-MS/MS）建立一种简单快速的测定人血浆中泽布替尼浓度的方法，并用于临床治疗药物监测。方法 以伊布替尼为内标，血浆样本经90%乙腈沉淀蛋白，采用Shim-pack GSP-HP C₁₈（50 mm×2.1 mm，3 μm）色谱柱，0.1%甲酸水-0.1%甲酸乙腈溶液为流动相，梯度洗脱，体积流量为0.3 mL·min-1，柱温40℃，进样量3 μL；质谱检测采用电喷雾离子源，正离子源，多反应监测模式，监测泽布替尼m/z 472.2→290.2（定量离子） ，m/z472.2→455.2（定性离子） ；伊布替尼m/z 441.2→138.1的浓度。28名受试者空腹口服泽布替尼160 mg，给药前30 min和给药后2 h收集血浆，按建立的UPLC-MS/MS法测定血浆中泽布替尼浓度，并将结果应用于临床治疗药物监测（ TDM）。结果 泽布替尼在1～1 000 ng·mL^-1内线性关系良好（ R²＝0.994 2） ，准确度在97.4%～104.3%，批内及批间精密度（ RSD）均≤4.8%，内标归一化基质效应在100.8%～102.5%，回收率为96.9%～100.5%。泽布替尼血浆样品在室温放置12 h，4℃放置12 h，进样器（8℃）放置24 h，反复冻融（ -20℃） 3次，-20℃放置30 d，-80℃放置30 d的情况下均稳定。28名患者泽布替尼平均稳态谷浓度为（ 3.25±1.75） ng·mL^-1，平均稳态峰浓度为（ 110.09±52.53） ng·mL^-1。TDM结果表明，泽布替尼谷浓度与患者肝功能指标密切相关。结论 建立的一种UPLC-MS/MS法测定人血浆中泽布替尼药物浓度，适用于泽布替尼TDM及个体化用药。

Objective To establish a simple and rapid ultra-performance liquid chromatography-tandem mass spectrometry (UPLCMS/MS) method for the determination of zanubrutinib concentration in human plasma and apply it to clinical therapeutic drug monitoring. Methods Ibrutinib was used as the internal standard. Plasma samples were precipitated with 90% acetonitrile. A Shimpack GSP-HP C₁₈ (50 mm×2.1 mm, 3 μm) column was used with a mobile phase of 0.1% formic acid water and 0.1% formic acid acetonitrile solution, gradient elution, a flow rate of 0.3 mL·min-1, column temperature of 40 ℃, and an injection volume of 3 μL. Mass spectrometry detection was performed using an electrospray ionization source, positive ion mode, and multiple reaction monitoring mode. The monitoring ions for zanubrutinib were m/z 472.2 → 290.2 (quantitative ion) and m/z 472.2 → 455.2 (qualitative ion), and the ion for ibrutinib was m/z 441.2 → 138.1. Twenty-eight subjects were given a single oral dose of 160 mg zanubrutinib after fasting. Blood samples were collected 30 minutes before administration and 2 hours after administration, and plasma was prepared. The plasma concentration of zanubrutinib was determined by the established UPLC-MS/MS method and the results were applied to clinical therapeutic drug monitoring (TDM). Results Zanubrutinib showed a good linear relationship within the range of 1—1 000 ng·mL^-1 (R² = 0.994 2), with accuracy ranging from 97.4% to 104.3%, and intra- and inter-batch precision (RSD) was ≤ 4.8%. The normalized matrix effect of the internal standard was between 100.8% and 102.5%, and the recovery rate was 96.9% to 100.5%. Zanubrutinib plasma samples were stable under conditions of room temperature for 12 hours, 4 ℃ for 12 hours, in the injector (8 ℃) for 24 hours, after three freeze-thaw cycles at -20 ℃, and at -20 ℃ for 30 days or at -80 ℃ for 30 days. The average steady-state trough concentration of zanubrutinib in 28 patients was (3.25 ±1.75) ng·mL^-1, and the average steady-state peak concentration was (110.09 ±52.53) ng·mL^-1. The TDM results indicated that the trough concentration of zanubrutinib was closely related to the liver function indicators of patients. Conclusion UPLC-MS/MS method was established for the determination of zanubrutinib concentration in human plasma, which is suitable for zanubrutinib therapeutic drug monitoring (TDM) and individualized medication.

R917

国家自然科学基金资助项目（82270175）；福建省自然科学基金资助项目（2021J01761）；福建省自然科学基金重点项目（2021J02040）