HPLC-MS/MS法测定Beagle犬血浆中盐酸格拉司琼的浓度

HPLC-MS/MS determination of granisetron hydrochloride in Beagle dog plasma

目的:建立HPLC-MS/MS法测定Beagle犬血浆中盐酸格拉司琼的浓度,用于盐酸格拉司琼的浓度测定及药代动力学研究。方法:采用8只健康Beagle犬分别交叉给予0.3 mg.kg-1盐酸格拉司琼片剂和盐酸格拉司琼贴剂,2次给药间隔为2周,不同时间点取血,血浆样品采用液-液萃取法处理。流动相为甲醇-乙腈-pH 4.5醋酸铵缓冲液(12.5 mmol.mL-1)(42∶18∶40),用Zorbax Eclipse XB-C18柱(5μm,150 mm×4.6 mm)色谱柱分离;以盐酸伪麻黄碱为内标。流速0.5 mL.min-1;进样量25μL,柱温为室温。质谱检测采用ESI正离子模式,扫描方式为多反应监测,扫描离子对m/z格拉斯琼313.2/138.1,伪麻黄碱166.1/148.1,用DAS2.1药代动力学软件计算药代动力学参数。结果:盐酸格拉司琼的线性范围为0.005～4 ng.mL-1,线性关系良好(r2=0.9986),最低定量限为0.005 ng.mL-1,回收率在95.3%～107.7%间,变异系数均小于7%。结论:本文所建立的HPLC-MS/MS法灵敏、准确、可靠,可用于犬血浆中盐酸格拉司琼的浓度测定及药代动力学研究。

Objective：To establish an HPLC-MS/MS method for determination of granisetron hydrochloride in Beagle dog plasma.Methods：A total of eight healthy Beagle dogs were randomly divided into two groups with a crossover design at the interval of 2 weeks and treated with granisetron hydrochloride tablets and patches at the same dose of 0.3 mg·kg^-1,respectively.The plasma samples were taken at different time points and processed by liquid-liquid extraction before analysis.The 25 μL processed sample was performed on a Zorbax Eclipse XB-C18 analytical column（5 μm,150 mm × 4.6 mm） using the organic phase[70% of methyl alcohol-30% of acetonitrile-12.5 mmol of ammonium acetate buffer（42∶ 18∶ 40）,pH=4.5]as the mobile phase,with a flow rate of 0.5 ml·min^-1.The mass spectrometer was operated under the positive ion mode and multiple reactions monitoring with the transitions of m/z 313.2/138.1 and m/z 166.1/148.1,respectively,to qualify granisetron and pseudoephedrine（as the internal standard）.The DAS 2.1 software was applied to analyze pharmacokinetic parameters.Results：The linear calibration curve was well obtained in the range of 0.005-4 ng·mL-1（r2=0.9986）,with the lower limit of quantification of 0.005 ng·mL-1.Recovery in the measurement of quality control samples was in the range of 95.3%-107.7% with variable coefficients both less than 7%.Conclusion：This method was simple,rapid and sensitive and could be used for determination of granisetron in Beagle dog plasma and pharmacokinetic study.