PMEA-Na在beagle犬血浆中的液相色谱/质谱/质谱联用法测定及其药代动力学

目的建立LC/MS/MS法测定犬血浆中PMEA-Na浓度,进行其药代动力学研究.方法血浆样品经甲醇沉淀蛋白后,采用多反应监测法测定其血药浓度.色谱柱为Xterra MS柱,流动相为甲醇水甲酸(25750.5),流速为 0.25 ml·min-1.Beagle犬分3个剂量组经静脉给药,给药剂量分别为 1.0、3.0 和 6.0 mg·kg-1.药代动力学参数通过DAS软件计算获得.结果PMEA-Na线性范围0.02～20 mg·L-1 (r=0.999);最低检测浓度为 20 μg·L-1,方法回收率为 97.1%～107.3%,日内日间变异分别小于 6.5%、10.8%.beagle 犬在 1.0, 3.0 与 6.0 mg·kg-1剂量下单次iv PMEA-Na后,测得其AUC分别为 2.3±0.5, 8.2±1.3 and 18.5±1.3 mg·L-1·h; t1/2 为 3.9±1.8, 8.4±1.5 and 8.9±0.6 h; CL为 0.44±0.09, 0.35±0.05 and 0.31±0.03 ml·h-1·kg-1.结论本方法专属性强,准确性好,可用于PMEA-Na血药浓度测定和药代动力学研究.

Determination of PMEA-Na in dog plasma by liquid chromatography and tandem mass spectrometry and its pharmacokinetic study

AIM: To established an HPLC/MS/MS method for the study of pharmcokinetics of PMEA-Na (the mono-sodium salts of 9-2-(phosphonomethoxy) ethyl] adenine) in beagle dogs. METHODS: PMEA-Na and internal standard 9-(3-phosphony-methoxypropyl) adenine were isolated from plasma by protein precipitation with methanol, and then analyzed adopting multiple reaction monitoring (MRM) mode. Using Xterra MS column, the mobile phases consisted of methanol:water:formic acid (25:75:0.5) at a flow rate of 0.25 ml·min-1. Beagle dogs received the intravenous dosage of PMEA-Na at 1.0, 3.0 and 6.0 mg·kg-1. Pharmacokinetic parameters were obtained from concentration-time curves by non-linear least-squares regression using the program DAS. RESULTS: The linear calibration curve was obtained in the concentration range of 0.02 to 20 mg·L-1 (r=0.999), and the limit of quantitition was 20 μg·L-1. The within-day and internal-day precisions (RSD) were less than 6.5% and 10.8%, respectively. The accuracy was 97.1%～107.3%. After a single dose studies in dogs the AUC were 2.3±0.5, 8.2±1.3 and 18.5±1.3 mg·L-1·h; the t1/2 were 3.9±1.8, 8.4±1.5 and 8.9±0.6 h; the CL were 0.44±0.09, 0.35±0.05 and 0.31±0.03 ml·h-1·kg-1 at the dose level of 1.0, 3.0 and 6.0 mg·kg-1 respectively. CONCLUSION: The analytical method is sensitive and specific for investigation the pharmacokintics of PMEA-Na in beagle dogs.