LC-MS/MS法测定大鼠血浆中阿霉素的药物浓度及应用

目的：建立液相色谱-串联质谱（LC-MS/MS）法测定大鼠血浆中阿霉素的血药浓度，并用于阿霉素载药纳米粒的体内药动学研究。方法色谱柱采用Shim-pack XR-ODS柱（2.0 mm＆#215;100 mm，2.2μm），仪器采用API 4000三重四极杆串联质谱仪，采用多反应监测（multiple reaction monitoring，MRM）定量离子对为m/z544.2→m/z396.9（阿霉素）和m/z357.3→m/z133.8（吡格列酮，内标）；甲醇沉淀蛋白法处理样品。结果阿霉素在2.0-2000μg/L范围内线性关系良好，定量下限为2.0μg/L，血浆中的内源性物质不干扰阿霉素的测定；日内和日间精密度RSD均小于±15.0%；阿霉素低、中、高3个QC浓度的提取回收率分别为（92.1±5.9）%、（82.8±2.9）%和（80.1±9.7）%，基质效应分别为（91.1±2.4）%、（82.1±1.7）%和（81.2±2.5）%。结论该方法适用于阿霉素纳米粒在大鼠体内的药物动力学研究。

Determination of Doxorubicin in rat plasma by LC-MS/MS and its appli-cation

Objective To develop a liquid chromatography-tandem mass spectrometry （LC-MS/MS） method for the de-termination of Doxorubicin in rat plasma and application of pharmacokinetics study. Methods The chromatographic separation was carried out on an Shim-pack XR-ODS （2.0 mm＆#237;100 mm, 2.2μm） and detected by an API 4000 mass spectrometer. The compounds were quantified by multiple-reaction monitoring （MRM） mode using transition mass of m/z 544.2→m/z396.9 for doxorubicin and m/z 357.3→m/z 133.8 for pioglitazone （internal standard）. Protein precipitation by methanol was used for the sample pretreatment. Results The calibration curves of Doxorubicin were linear over the range 2-2000μg/L with the lower limit of quantication of 2.0μg/L. Determination of Doxorubicin was not interfered by endogenous substance in plasma. The intra-day and inter-day precisions of QC samples were within±15.0% （RSD%）. The extraction recoveries and matrix effects of QC samples were （92.1±5.9）%, （82.8±2.9）%, and （80.1±9.7）%; （91.1±2.4）%, （82.1±1.7）%, and （81.2±2.5）%, respectively. Conclusion This method can be applied to pharmacokinetic studies of Doxorubicin loaded nanoparticles in rats.