LC-MS法测定Beagle犬血浆中多西他赛的浓度

Determination of docetaxel concentration in Beagle dog plasma by LC-MS

目的建立快速检测Beagle犬血浆中多西他赛浓度的液相色谱-质谱联用方法。方法取100μL犬血浆,以200μL含有50μg·L^(-1)内标(紫杉醇)的乙腈溶液沉淀蛋白,10 000 r·min^(-1)5 min离心2次,取上清液进样10μL。色谱柱:Agilent Eclipse Plus C_(18)(150 mm×2.1 mm,5μm),柱温:30℃,流动相:甲醇-水相梯度洗脱,流速0.2 mL·min^(-1)。ESI源正离子SRM模式,定量的子离子:多西他赛[M+H]^+m/z:830.3→549.3(26 eV),紫杉醇[M+H]^+m/z:876.3→308.1(27 eV)。结果多西他赛的线性范围为5～1 000μg·L^(-1),标准曲线的相关系数r≥0.996 5,提取回收率为89.9%～114.8%,方法回收率为94.0%～97.5%,批内变异系数<9.0%,批间变异系数<10.8%。结论本方法简便、迅速、灵敏,方法学各项指标均符合要求,适用于犬血浆中多西他赛浓度的检测。

AIM To establish an LC-MS method for the quick determination of docetaxel concentration in plasma of Beagle dog.METHODS The 200 μL acetonitrile that contained 50 μg·L^-1 paclitaxel (internal standard)was added to 100 μL dog plasma for protein precipitation.After centrifuging for 5 min twice at a speed of 10 000 r ·min^-1,10 μL supernatant was injected to LC-MS.Agilent Eclipse Plus C18 column (150 mm ×2.1 mm,5 μm)was adopted and the temperature was set at 30℃.Methanol and water was used as mobile phase and the follow rate was set as 0.2 mL·min ^-1.Under the ESI positive ion SRM mode,the daughter ion [M +H ]^+m/z 549.3 (parent ion [M +H ]^+m/z 830.3 )and 308.1 (parent ion [M +H ]^+m/z 876.3 )were adopted for quantitative determination for docetaxel and paclitaxel,and with the collision energy of 26 eV and 27 eV,respectively. RESULTS The linear range was within 5 -1 000 μg·L^-1 and the correlation coefficient (r)was more than 0.996 5.The extraction recovery was within 89.9%-114.8%and the assay recovery was within 94.0%-97.5%.The coefficients of variability of intra-batch precision and inter-batch were less than 9.0%and 10.8%,respectively. CONCLUSION The method is convenient,quick and sensitive and all the items are validated for the determination of docetaxel concentration in dog plasma.