HPLC-MS/MS法测定白血病患者血浆奥雷巴替尼的方法学研究及临床应用

Methodological study and clinical application of HPLC-MS/MS for the determination of olverembatinib in the plasma of leukemia patients

目的建立测定白血病患者血浆奥雷巴替尼浓度的高效液相色谱串联质谱法,并应用于临床治疗药物监测,为临床合理用药提供可靠依据。方法以帕纳替尼-d8为内标,用甲醇沉淀血浆蛋白和萃取奥雷巴替尼。色谱柱为Welch Ultimate XB-C18柱(50mm×4.6 mm,5μm),柱温为60℃,流动相为水溶液(含0.1%甲酸+2 mmol/L乙酸铵)-甲醇溶液(含0.1%甲酸),流速为0.8 mL/min,梯度洗脱;采用电喷雾正离子模式,多反应监测扫描,奥雷巴替尼定量离子对m/z 533.3→260.1和定性离子对m/z 533.3→433.3,内标离子对m/z 541.1→260.2。对40例服用奥雷巴替尼的白血病患者血浆样本进行浓度监测,运用IBM SPSS Statistics 27.0和OriginPro 2021软件对结果数据进行统计分析。结果奥雷巴替尼线性范围为1~250 ng/mL(r=0.9980),定量下限为1 ng/mL,提取回收率为100.28%~101.27%,日内精密度RSD为1.15%~3.87%,日间精密度RSD为2.32%~3.68%。结论该方法操作简便、特异性强、灵敏度高,可用于测定白血病患者奥雷巴替尼血药浓度。

Objective To establish a high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)method for determining the plasma concentration of olverembatinib in leukemia patients,apply it to clinical drug monitoring,and provide reliable basis for rational drug use in clinical practice.Methods Ponatinib-d8 was used as an internal standard,and methanol was used to precipitate plasma proteins and extract olverembatinib.The chromatographic column was Welch Ultimate XB-C18 cloumn(50 mm×4.6 mm,5μm),with a column temperature of 60°C.The mobile phase consisted of an aqueous solution(containing 0.1%formic acid+2 mmol/L ammonium acetate)-methanol solution(containing 0.1%formic acid),with a flow rate of 0.8 mL/min and gradient elution.Electrospray positive ion mode was used,with multiple reaction monitoring scanning.The quantitative ion pair of olverembatinib was m/z 533.3→260.1,the qualitative ion pair was m/z 533.3→433.3,and the internal standard ion pair was m/z 541.1→260.2.The plasma samples of 40 leukemia patients taking olverembatinib were monitored and analyzed for concentration,and IBM SPSS Statistics 27.0 and OriginPro 2021 softwares were used for statistical analysis of the results.Results The linear range of olverembatinib was 1-250 ng/mL(r=0.9980),the lower limit of quantification was 1 ng/mL,the extraction recovery rate was 100.28%~101.27%,the intra-day precision RSD was 1.15%~3.87%,and the inter-day precision RSD was 2.32%~3.68%.Conclusion This method is easy to operate,highly specific and sensitive,and can be used to determine the blood concentration of olverembatinib in leukemia patients.