LC-MS/MS法测定人血浆中白消安的浓度

目的: 建立液相色谱-串联质谱(LC-MS/MS)法检测人血浆中白消安的浓度。 方法: 取血浆样品100 μL,采用含内标1,5-戊二醇二甲磺酸酯的乙腈沉淀蛋白,离心后取上清液进样分析。色谱柱为Agilent Eclipse XDB-C₁₈(100 mm×2.1 mm,3.5 μm),柱温40 ℃,流动相为2 mmol·L^-1甲酸铵(含0.1%甲酸)-乙腈(60∶40,v/v),流速为0.20 mL·min^-1;采用电喷雾化离子源(ESI),多离子反应模式(MRM)检测,白消安和内标的离子对分别为m/z 264.2→151.1和m/z 278.3→69.2。 结果: 白消安的线性范围为4.875~3120 μg·L^-1,检测下限浓度为4.875 μg·L^-1,低、中、高3个浓度质控样品日内和日间RSD均<15%,平均提取回收率为102.0%~113.7%。 结论: 该方法样本处理简便快速,灵敏度较高,所测得结果准确可靠,已成功应用于白消安人体内药动学研究,适用于其血药浓度常规监测。

Objective: To establish an LC-MS/MS method for determining concentration of busulfan in human plasma.Methods: Totally 100 μL of plasma samples containing busulfan was precipitated by acetonitrile containing 1,5-pentanediol dimethylsulfonate as the internal standard,the samples were centrifuged and the supernatant was injected into LC-MS/MS.HPLC separation was performed on an Agilent Eclipse XDB-C₁₈ analytical column (100 mm×2.1 mm,3.5 μm) with a column temperature of 40 ℃.Samples were eluted using a mobile phase composed of 2 mmol·L^-1 ammonium formate aqueous (0.1% formic acid aqueous)-acetonitrile (60∶40,v/v) at a flow rate of 0.2 mL·min^-1.ESI was used which was performed in the multiple reaction monitoring mode [MRM using target ions m/z 264.2→151.1 (busulfan),m/z 278.3→69.2 (IS)].Results: The linear range of busulfan was 4.875-3120 μg·L^-1 with LLOQ of 4.875 μg·L^-1.The intra-and inter-day RSDs were all below 15% at low,moderate and high concentrations.The mean extract recoveries were 102.0%-113.7%.Conclusion: The method established is rapid,sensitive,accurate and reliable for the determination of busulfan in human plasma.This method has been successfully applied in the study on busulfan pharmacokinetics in human and can be used for the conventional monitoring of the plasma concentration of busulfan.

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