基于LC-MS/MS法测定人血浆仑伐替尼浓度及临床应用分析

Determination of blood concentration of lenvatinib in human plasma by LC-MS/MS and clinical application analysis

关键词:  仑伐替尼  肝细胞癌  血药浓度  治疗药物监测

Key words:  lenvatinib  hepatocellular carcinoma  blood drug concentration  therapeutic drug monitoring

摘要:

目的 建立测定血浆中仑伐替尼浓度的液相色谱串联质谱(HPLC-MS/MS)方法,并用于临床用药监测。方法 利用Waters HSS T3色谱柱,以仑伐替尼-d5同位素为内标,以去离子水(含0.1%甲酸)和乙腈(含0.1%甲酸)为流动相进行梯度洗脱,流速0.4 mL·min-1,柱温40℃,进样量2 uL；采用电喷雾正离子(ESI+)模式多重反应监测(MRM)进行检测。选择本院普外科接收受仑伐替尼治疗的肝细胞癌(HCC)患者,测定其仑伐替尼血药浓度。结果 构建的仑伐替尼检测方法在1.15~410.38 ng·mL-1范围内线性关系良好(r>0.999),定量下限为1.15 ng·mL-1 ,可以满足临床应用。日内、日间准确度为96.97%~102.68%,精密度RSD均小于15%；提取回收率为94.21%~100.48%；内标归一化基质效应为99.75%~108.84%,RSD均小于15%；稳定性试验准确度在97.11%~103.78%,RSD均小于15%；方法学验证均满足生物样品分析要求。完成仑伐替尼谷浓度检测54人次,浓度为 2.387-67.453 ng·mL-1,平均浓度为21.60 ng·mL-1,存在明显个体差异。结论 构建的仑伐替尼浓度LC-MS/MS测定法准确、高效,可用于仑伐替尼用药监测及药动学研究。

Abstract:

OBJECTIVE To establish a HPLC-MS/MS method for determining the concentration of lenvatinib in plasma and apply it to clinical drug monitoring. METHODS The determination was performed using Waters HSS T3 chromatographic column with lenvatinib-d5 isotope as the internal standard, gradient elution was performed with deionized water (containing 0.1% formic acid) and acetonitrile (containing 0.1% formic acid) as the mobile phases at a flow rate of 0.4 mL·min-1, column temperature of 40 ℃, and injection volume of 2 uL. Multiple reaction monitoring (MRM) with electrospray positive ion (ESI+) mode was used for detection.The patients with hepatocellular carcinoma (HCC) receiving treatment with lenvatinib in our general surgery department were selected and the blood concentration of lenvatinib were measured. RESULTS The constructed detection method for lenvatinib has a good linear relationship within the range of 1.15-410.38 ng·mL-1 (r>0.999), with a lower limit of quantification of 1.15 ng·mL-1, which can meet clinical applications. The intra day and inter day accuracy ranges from 96.97% to 102.68%, and the RSD of precision was less than 15%. The extraction recovery rate ranges from 94.21% to 100.48%,and the normalized matrix effect of the internal standard ranges from 99.75% to 108.84%, with RSD less than 15%. The accuracy of stability testing ranges from 97.11% to 103.78%, with RSD less than 15%. The methodological validation meets the requirements for biological sample analysis. 54 individuals were tested for the concentration of lenvatinib, with concentrations ranging from 2.387-67.453 ng·mL-1 and an average concentration of 21.60 ng·mL-1, showing significant individual differences. CONCLUSION The LC-MS/MS method for determining the concentration of lenvatinib constructed is accurate and efficient, and can be used for monitoring the use of lenvatinib and pharmacokinetic studies.