周利娟 武正华 汪硕闻 缪文清 包婺平 包爱华 范国荣

摘 要 目的：建立测定人血浆中安罗替尼质量浓度的方法并应用于临床。方法：以醋酸铵为盐析助剂，以乙腈为溶剂，采用盐析辅助液液萃取对血浆样品进行前处理;以伏立康唑为内标，采用液相色谱-串联质谱法（LC-MS/MS）测定。色谱柱为Waters X Bridge C18，流动相为0.2%甲酸水溶液-乙腈（梯度洗脱），流速为1 mL/min，柱温为40 ℃，进样量为10 μL，分流比为3 ∶ 7;离子源为电喷雾离子源，以多反应监测模式进行正离子扫描，用于定量分析的离子对分别为m/z 408.3→339.3（安罗替尼）、m/z 350.2→281.3（内标）。结果：安罗替尼检测质量浓度的线性范围为0.2～200 ng/mL（R2=0.996 7）;定量下限为0.2 ng/mL;日内、日间精密度试验的RSD均小于12%（n=6或n=3）;准确度为90.92%～108.00%（n=6或n=3）;平均提取回收率为87.51%～100.00%（RSD<8%，n=6）;平均基质效应为96.66%～99.93%（RSD<5%，n=6）。3例使用安罗替尼治疗的非小细胞肺癌患者体内的血药浓度为8.74～65.60 ng/mL。结论：该方法操作简单、准确度高、专属性强，可用于非小细胞癌患者体内安罗替尼的血药浓度监测。

关键词 安罗替尼;液相色谱-串联质谱技术;盐析辅助液液萃取;治疗药物监测

ABSTRACT   OBJECTIVE： To establish a method for concentration determination of anlotinib in human plasma and apply it in the clinic. METHODS： The plasma samples were pretreated by salting-out assisted with liquid-liquid extraction with ammonium acetate as salting out assistant and acetonitrile as solvent. Using voriconazole as internal standard， LC-MS/MS method was adopted. The separation was performed on Waters X Bridge C18 column with mobile phase consisting of 0.2% formic acid solution- acetonitrile （gradient elution） at the flow rate of 1 mL/min. The column temperature was set at 40 ℃， and sample size was 10 μL. The split ratio was 3 ∶ 7. The electrospray ion source and multiple reaction monitoring mode were used for the analysis. The ion pair of anlotinib and internal standard under positive ion mode were m/z 408.3→339.3 and m/z 350.2→281.3， respectively. RESULTS： Anlotinib showed a good linear relationship in the concentration range of 0.2-200 ng/mL （R2>0.996 7）. The lowest limit of quantitation was 0.2 ng/mL. Intra-day and inter-day RSDs were no more than 12% （n=6 or n=3）. Accuracies were 90.92%-108.00% （n=6 or n=3）. The average extraction recoveries were 87.51%-100.00%（RSD<8%，n=6）. The average matrix effects were 96.66%-99.93%（RSD<5%，n=6）. The plasma concentration of 3 patients with NSCLC treated with anlotinib was 8.74-65.60 ng/mL. CONCLUSIONS： The method is simple， accurate and specific， and is suitable for the plasma concentration monitoring of anlotinib in NSCLC patients.

KEYWORDS   Anlotinib; LC-MS/MS; Salting-out assisted liquid-liquid extraction; Therapeutic drug monitoring

鹽酸安罗替尼是我国自主研发的一种新型口服酪氨酸激酶抑制剂，能够有效抑制血管内皮生长因子受体、血小板衍生生长因子受体、纤维母细胞生长因子受体、干细胞因子受体（c-Kit）等激酶，具有抗肿瘤血管生成、抑制肿瘤生长的双重作用[1-2]。基于ALTER 0303研究（安罗替尼治疗晚期非小细胞肺癌患者的Ⅲ期临床研究）的良好结果，国家食品药品监督管理局于2018年5月8日正式批准安罗替尼单药用于晚期非小细胞肺癌患者的三线及以上治疗。有研究表明，安罗替尼的药动学特征与其他酪氨酸激酶抑制剂类似，即经肝药酶代谢，其血浆蛋白结合率较高、消除半衰期较长，长期连续使用后药物在体……