LC-MS/MS法测定Beagle犬血浆中阿戈美拉汀的质量浓度

Determination of agomelatine in Beagle dogs plasma by LC-MS/MS

目的建立LC-MS/MS的方法测定Beagle犬血浆中阿戈美拉汀的质量浓度,并应用此方法研究阿戈美拉汀片在Beagle犬体内的药代动力学行为及其相对生物利用度。方法采用：Agela C18色谱柱（150 mm×4.6 mm,5μm I.D）,以乙腈-浓度5 mmol·L-1醋酸铵水溶液-甲酸（体积比为58.0∶42.0∶0.1）为流动相,采用沉淀蛋白法,以多反应离子监测（multiple reaction monitoring,MRM）扫描方法进行检测,测定经口给予阿戈美拉汀片后Beagle犬血浆中阿戈美拉汀的质量浓度。结果 Beagle犬血浆中阿戈美拉汀在质量浓度0.50~100μg·L-1内与峰面积呈良好线性关系,相关系数r为0.999 0;日内和日间精密度RSD≤8.6%;阿戈美拉汀的平均提取回收率为90.2%~93.3%,基质效应为104.9%~106.8%;阿戈美拉汀在Beagle犬血浆中主要药动学参数t1/2为（4.2±1.5）h,ρmax为（149±85）μg·L-1,AUC0-24为（293±78）μg·h·L-1,AUC0-∞为（300±78）μg·h·L-1。结论该方法适用于阿戈美拉汀在Beagle犬体内的药代动力学及相对生物利用度的研究。

Objective To develop a liquid chromatography-tandem mass spectrometry（ LC-MS/MS） method for the determination of agomelatine in Beagle plasma,and to study the pharmacokinetic characteristic and bioequivalence of agomelatine in Beagles plasma. Methods The chromatographic separation was accomplished on Agela C18 analytical column（ 150 mm×4.6 mm I. D,5 μm） with the mobile phase consisting of acetonitrile： 5 mmol·L- 1ammonium acetate solution： formic acid（ V ∶ V ∶ V = 58. 0∶ 42. 0∶0.1）. Protein precipitation was used for the sample pretreatment. The plasma concentration of agomelatine was detected by multiple reaction monitoring after the Beagle dogs were given agomelatine tablets. Results The linear calibration curve of agomelatine was obtained in the concentration range of 0. 50- 100 μg·L- 1,r = 0. 999 0; The intraand inter-day precision was both less than 8. 6%. The average recoveries were between 90. 2% and 93. 3%.The matrix effects were between 104. 9% and 106. 8%. The main pharmacokinetic parameters were as follows： t1 /2（ 4. 2 ± 1. 5） h,ρmax（ 149 ± 85） μg·L- 1,AUC0- 24（ 293 ± 78） μg·h·L- 1,AUC0- ∞（ 300 ±78） μg·h·L- 1,respectively. Conclusions This simple and rapid LC-MS/MS method is successfully applied to pharmacokinetic studies and relative bioavailability of agomelatine in Beagle dogs plasma.