液相色谱-串联质谱法研究伏立康唑在犬血浆的毒代动力学

Determination of Voriconazole in Dog Plasma by LC-MS/MS and Its Toxicokinetics

目的建立液相色谱-串联质谱法测定犬血浆中伏立康唑的浓度并研究其毒性代谢动力学。方法犬血浆样本以乙腈沉淀蛋白后,选用Agilent Poroshell 120 EC-C18色谱柱(2.1 mm×50 mm, 2.7 μm),流动相为0.1%甲酸水与乙腈,流速为0.3 mL·min-1,柱温为40 ℃,进样量为10 μL,分析时间为5 min;选用Agilent 6460三重四极杆串联质谱仪,离子化方式:电喷雾-正离子(API-ES),干燥气:N2,干燥气流速11 L·min-1,干燥气压力0.31 MPa,干燥气温度为350 ℃,毛细管电压4 kV。监测方式:MRM,伏立康唑监测离子对m/z 335.1/281.3,氟康唑监测离子对m/z 307.1/220.0。30只Beagle犬,静脉输注伏立康唑低、中、高3个剂量组(1、3、6 mg·kg-1·d-1),每天静脉输注给药1次,连续给药12周。结果本分析方法专属性良好;伏立康唑在10~10 000 ng·mL-1内线性关系符合要求;准确度均在85%~115%,精密度RSD在15%内。伏立康唑在比格犬体内雌性暴露量明显高于雄性,当给药1~6 mg·kg-1·d-1时,伏立康唑在比格犬体内暴露量随着剂量增加而增大,但二者之间不太符合线性关系,药物基本无蓄积。结论本法经方法学验证,适用于犬血浆中伏立康唑浓度的测定,可用于伏立康唑犬体内毒代动力学研究。

OBJECTIVE To establish a method for the determination of voriconazole in dog plasma and investigate its toxicokinetics. METHODS After protein precipitation with acetonitrile, voriconazole and fluconazole were separated on an Agilent Poroshell120 EC-C18 column (2.1 mm×50 mm, 2.7 μm), with acetonitrile and water (0.1% FA) as the mobile phase at a flow rate of 0.3 mL·min-1. Detection was carried out by the electrospray positive ionization mass spectrometry in the multiple reacion monitoring (MRM) mode. The MRM transitions of m/z 335.1→281.3 and m/z 307.1→220.0 were used to quantify voriconazole and fluconazole, respectively. Thirty Beagle dogs received intravenous infusion of voriconazole at low, medium and high doses (1, 3, 6 mg·kg-1·d-1) once a day for 12 weeks. RESULTS The calibration curve was linear over 10-10 000 ng·mL-1 . RSD was less than 15%, and the accuracy was within the range of 85%-115%.The exposure of voriconazole to females was significantly higher than that of males in Beagle dogs. When the dose was 1-6 mg·kg-1·d-1, the exposure of voriconazole to beagle dogs increased with the increase of dose, and the drug did not accumulate. CONCLUSION The method can be applied to the determination of voriconazole in dog plasma, and is suitable to the toxicokinetics study of voriconazole.