高效液相色谱-串联质谱法测定大鼠血浆中抗肿瘤化合物WJD-A-1及其药动学研究

Pharmacokinetics of WJD-A-1 and WJD-A-1 in rat plasma determined by HPLC-MS/MS

目的:建立大鼠血浆中抗肿瘤化合物WJD-A-1的高效液相色谱-串联质谱(HPLC-MS/MS)测定方法,并研究静脉注射WJD-A-1后在大鼠体内的药动学特征。方法:以苯海拉明为内标,大鼠血浆样品经甲醇-乙腈处理,用色谱柱Waters ACQUITY UPLC■ BEH C18(2.1 mm×50 mm, 1.7μm)进行分离,以甲醇-0.1%甲酸(80∶20)为流动相,采用电喷雾离子源(ESI)正离子模式及多重反应监测模式,检测离子对分别为WJD-A-1(m/z 480.6→334.6,480.6→164.3)以及苯海拉明(m/z 256.0→166.8)。将建立的检测方法应用于测定大鼠静脉注射化合物WJD-A-1后的血浆浓度。结果:大鼠血浆中化合物WJD-A-1在5.40~5.40×103 ng·mL-1浓度范围内线性关系良好;低、中、高3个质控浓度下的日内、日间精密度均小于10%,质控样品的准确度为97.45%~106.80%;提取回收率为95.40%~96.17%,无明显基质效应且稳定性良好。大鼠静脉给与化合物WJD-A-1三个剂量75,150,300μg·kg^-1后,血浆中WJD-A-1的半衰期t1/2分别为(0.10±0.05)h、(0.24±0.02)h、(0.51±0.06)h;其浓度-时间曲线下的面积AUC(0-t)分别为(150.60±21.69)μg·h·L^-1、(883.07±135.68)μg·h·L^-1、(3 311.59±418.15)μg·h·L^-1。结论:该检测方法简单、快速、灵敏、准确,适用于抗肿瘤化合物WJD-A-1在大鼠血浆内的浓度测定及其药动学研究。药动学参数表明大鼠静脉注射该化合物后体内消除较快,且该化合物在大鼠体内的暴露量与给药剂量成正比。

OBJECTIVE To establish an HPLC-MS/MS method for the determination of antineoplastic compounds, WJD-A-1 in rat plasma and to study the pharmacokinetic characteristics of WJD-A-1 in rats after intravenous injection. METHODS The plasma samples of rats were treated with methanol acetonitrile and separated by chromatographic column Waters ACQUITY UPLC BEH C18(2.1 mm×50 mm, 1.7 μm). Methanol-0.1% formic acid(80∶20) was used as mobile phase, and electrospray ionization ion source(ESI) positive mode and multiple reaction monitoring mode were used to detect ion pairs respectively WJD-A-1(m/z 480.6→334.6,480.6→164.3) and internal standard diphenhydramine(m/z 256.0→166.8). The established method was applied to determine the plasma concentration of compound WJD-A-1 after intravenous injection in rats. RESULTS The linear relationship of WJD-A-1 in rat plasma was good in the concentration range of 5.40-5.40×103 ng·mL^-1. The intra-day and inter-day precision RSD of low, medium and high quality control concentrations were all less than 10%, and the accuracy of quality control samples was 97.45%-106.80%. The recovery of WJD-A-1 was 95.40%-96.17%. There was no obvious matrix effect and stability was good. The half-life t1/2 of WJD-A-1 in plasma was(0.10±0.05)h、(0.24±0.02)h、(0.51±0.06)h after intravenous administration of compound WJD-A-1 75, 150, and 300 μg·kg^-1 in rats. The area under the concentration-time curve, AUC(0-t) was(150.60±21.69)μg·h·L^-1、(883.07±135.68)μg·h·L^-1、(3 311.59±418.15)μg·h·L^-1. CONCLUSION The method is simple, rapid, sensitive and accurate. It is suitable for the determination of WJD-A-1 in rat plasma and research of pharmacokinetics. Pharmacokinetic parameters showed that the compound was eliminated quickly in rats after intravenous injection, and the exposure of the compound in rats was proportional to the dosage of the drug.