固相萃取-UFLC-MS/MS法快速测定血浆中多柔比星浓度

Rapid determination of doxorubicin in plasma by solid-phase extraction and ultrafast liquid chromatography tandem mass spectrometry

目的建立一种快速、灵敏、准确的测定血浆中多柔比星浓度的固相萃取-超快速液相色谱-串联质谱联用(UFLC-MS/MS)方法。方法血浆样品经4%磷酸溶液提取后采用PRiME MCX萃取小柱净化,在Shim-pack XR-ODSⅡ柱(100 mm×2.0 mm,1.7μm)上,以含0.1%甲酸的水溶液(A相)和含0.1%甲酸的乙腈溶液(B相)为流动相进行梯度洗脱,以柔红霉素为内标,采用电喷雾离子源(ESI)在多反应监测(MRM)正离子模式下检测,多柔比星和内标的定量离子对分别为m/z544.3→397.0和m/z 528.4→320.9,定性离子对分别为m/z 544.3→379.2和m/z 528.4→363.0。结果多柔比星在质量浓度0.2~200.0μg·L^(-1)时具有良好的线性,定量检测下限(LLOQ)为0.2μg·L^(-1),其批内和批间RSD分别<10.7%和11.9%。相比Oasis HLB萃取小柱,PRiME MCX小柱的绝对基质效应平均值和提取平均回收率可分别提高14.3%和25.0%。结论采用PRiME MCX小柱,不仅可以降低基质效应提高回收率,而且可以缩短分析时间,简化操作程序。所建立的方法可用于患者血浆中痕量多柔比星的快速准确测定。

AIM To establish a fast,sensitive and accurate method for the determination of doxorubicin in plasma sample by solid-phase extraction and ultrafast liquid chromatography tandem mass spectrometry(UFLC-MS/MS).METHODS The separation was carried out on a Shim-pack XR-ODSⅡcolumn(100 mm×2.0 mm,1.7μm)using 0.1%formic acid-water solution and 0.1%formic acid-acetonitrile as mobile phase in gradient elution after plasma samples were extracted by 4%phosphoric acid solution and cleaned by PRiME MCX solid-phase extraction cartridge.Detection was performed by an electrospray ionization(ESI)in the positive multipile reaction monitoring(MRM)mode using daunorubicin as an internal standard(IS).The ion pairs m/z 544.3→397.0 and m/z 528.4→320.9 were selected for quantification for doxorubicin and IS,the ion pairs m/z 544.3→379.2 and m/z 528.4→363.0 were selected for qualitative analysis,respectively.RESULTS Within-batch and between-batch relative standard deviations were less than 10.7%and 11.9%,respectively.The linear range was 0.2-200.0μg·L^(-1) for doxorubicin.The lower limit of quantification(LLOQ)in plasma sample was 0.2μg·L^(-1).Compared to Oasis HLB extraction cartridge,the average values of absolute matrix effect and average extraction recovery of PRiME MCX cartridge increased by 14.3%and 25.0%,respectively.CONCLUSION The use of PRiME MCX cartridge can not only reduce absolute matrix effect and improve recovery,but also shorten the analysis time,simplify the operation process.The established method can be applied to rapid and accurate determination of trace doxorubicin in the patient's plasma sample.