目的 采用超高效液相色譜-串聯質譜（ UPLC-MS/MS）建立一種簡單快速的測定人血漿中澤布替尼濃度的方法，并用于臨床治療藥物監(jiān)測。方法 以伊布替尼為內標，血漿樣本經90%乙腈沉淀蛋白，采用Shim-pack GSP-HP C₁₈（50 mm×2.1 mm，3 μm）色譜柱，0.1%甲酸水-0.1%甲酸乙腈溶液為流動相，梯度洗脫，體積流量為0.3 mL·min-1，柱溫40℃，進樣量3 μL；質譜檢測采用電噴霧離子源，正離子源，多反應監(jiān)測模式，監(jiān)測澤布替尼m/z 472.2→290.2（定量離子） ，m/z472.2→455.2（定性離子） ；伊布替尼m/z 441.2→138.1的濃度。28名受試者空腹口服澤布替尼160 mg，給藥前30 min和給藥后2 h收集血漿，按建立的UPLC-MS/MS法測定血漿中澤布替尼濃度，并將結果應用于臨床治療藥物監(jiān)測（ TDM）。結果 澤布替尼在1～1 000 ng·mL^-1內線性關系良好（ R²＝0.994 2） ，準確度在97.4%～104.3%，批內及批間精密度（ RSD）均≤4.8%，內標歸一化基質效應在100.8%～102.5%，回收率為96.9%～100.5%。澤布替尼血漿樣品在室溫放置12 h，4℃放置12 h，進樣器（8℃）放置24 h，反復凍融（ -20℃） 3次，-20℃放置30 d，-80℃放置30 d的情況下均穩(wěn)定。28名患者澤布替尼平均穩(wěn)態(tài)谷濃度為（ 3.25±1.75） ng·mL^-1，平均穩(wěn)態(tài)峰濃度為（ 110.09±52.53） ng·mL^-1。TDM結果表明，澤布替尼谷濃度與患者肝功能指標密切相關。結論 建立的一種UPLC-MS/MS法測定人血漿中澤布替尼藥物濃度，適用于澤布替尼TDM及個體化用藥。

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

國家自然科學基金資助項目（82270175）；福建省自然科學基金資助項目（2021J01761）；福建省自然科學基金重點項目（2021J02040）