目的 建立一种简便、快速、灵敏的测定大鼠多柔比星血药浓度的超高效液相-质谱联用（UPLC-MS/MS）法，并将其应用于注射用盐酸多柔比星大鼠体内毒代动力学实验。方法 采用ACQUITY UPLC^® BEH C₁₈（50 mm×2.1 mm，1.7 μm）色谱柱，流动相为0.1%甲酸（含2 mmol/L甲酸铵）水溶液-乙腈，梯度洗脱。体积流量为0.4 mL/min，进样量为10 μL。采用电喷雾离子源（ESI），多反应监测（MRM）方式扫描，以正离子方式进行检测，蛋白沉淀法提取样品。用于定量分析的离子对分别为多柔比星m/z 544.43→m/z 397.08，内标地西泮m/z 285.02→154.40。SD大鼠30只，按体质量随机分为3组，分别单次iv 52.2、61.4、72.3 mg/m²盐酸多柔比星后测定血药浓度，并用DAS 3.1.4软件计算毒代参数。结果 血浆中内源性物质不干扰待测物和内标的测定，多柔比星在0.5～100 ng/mL范围内线性关系良好，定量下限为0.5 ng/mL。多柔比星在0.5、1、20、80 ng/mL 4个浓度的批内批间精密度RSD值为3.21%～12.79%。多柔比星在1、80 ng/mL的提取回收率和基质效应分别为102.00%～103.75%和79.27%～89.34%。SD大鼠分别单次iv给予注射用盐酸多柔比星52.2、61.4、72.3 mg/m²后，多柔比星在大鼠体内的AUC_0-t分别为（2 318.78±282.65）、（3 203.11±829.41）和（3 326.96±546.04） ng·h/mL，C_0.083h分别为（1 720.50±851.19）、（3 363.00±1 458.84）和（2 156.50±919.90）ng/mL。结论 建立的UPLC-MS/MS分析方法灵敏度高、样品处理方法简单、样品分析时间短，可以应用于大鼠多柔比星毒代动力学试验中。

Objective A simple, rapid, and sensitive ultra high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method has been developed to detect doxorubicin in rat plasma. The method was also validated for its application in toxicokinetics. Methods The separation was carried out on ACQUITY UPLC^® BEH C₁₈ column (50 mm×2.1 mm, 1.7 μm). The mobile phase consisted of 0.1% formic acid with 2 mmol/L ammonium formate and acetonitrile with gradient elution. The flow rate was 0.4 mL/min, and volume of injection was 10 μL. Electrospray ionization (ESI) source and multiple-reaction monitoring (MRM) was performed in the positive ion mode. The samples were prepared by protein precipitation. The ion pairs of m/z 544.43→m/z 397.08 and m/z 285.02→m/z 154.40 were used to doxorubicin and internal standard diazepam. A total of 30 SD rats were randomly divided into three groups according to body weight. The concentration of doxorubicin in rat plasma after intravenous administration of doxorubicin hydrochloride injection of 52.5, 61.4, and 72.3 mg/m² were determined, and the toxicokinetic parameters were calculated by using DAS 3.1.4. Results Endogenous substance in plasma had no effect on results. The linear ranges of doxorubicin were 0.5 — 100 ng/mL, with the lower limit of quantification of 0.5 ng/mL. Under concentrations of 0.5, 1, 20, and 80 ng/mL, the RSD value of intra and inter-day were 3.21%—12.79%. The extraction recovery and matrix effect of doxorubicin at 1 and 80 ng/mL were 102.00% — 103.75% and 79.27% — 89.34%, respectively. After intravenous administration of doxorubicinhydrochloride injection of 52.5, 61.4, and 72.3 mg/m², the toxicokinetic parameters were as follows: AUC_0-t (2 318.78 ±282.65), (3 203.11 ±829.41) and (3 326.96 ±546.04) ng·h/mL; C_0.083h (1 720.50 ±851.19), (3 363.00 ±1 458.84) and (2 156.50 ±919.90) ng/mL, separately. Conclusion The established UPLC-MS/MS method has high sensitivity, simple sample preparation method and short sample analysis time. It can be applied to the toxicokinetic test of doxorubicin in rats.

R969.1

国家科技重大专项（2015ZX09501004）