猪血浆中格拉司琼质量浓度的LC-MS/MS测定方法及应用

Study and application on the determination of granisetron in pig plasma by LC-MS / MS

目的建立猪血浆中格拉司琼质量浓度液相色谱-串联质谱（LC-MS／MS）检测方法，考察格拉司琼经皮给药后药物动力学行为。方法色谱柱：DiamonsilC18柱（150mm×4．6mm，5μm），流动相：甲醇-5mmol·L^-1醋酸铵-甲酸（体积比为45：55：0．1），采用液液萃取法，以多反应监测（multiple reaction monitoring，MRM）扫描方式，测定巴马小型猪经皮给予格拉司琼贴剂后血浆中药物的质量浓度。结果猪血浆中格拉司琼质量浓度在0．020～20．0μg·L^-1内线性关系良好，r=0．9959；日内和日间精密度RSD≤13．4％；格拉司琼的平均提取回收率为63．8％～66．7％（n=6），基质效应为101．4％～105．8％（n=6）；格拉司琼在巴马小型猪血浆中主要药物动力学参数：t1/2为（18．0±1．4）h，ρmax为（15．3±4．0）μg·L^-1，MRT为（91．9±4．9）h，Ke为（0．04±0.00）h^-1，AUC0.216为（1．224±0．35）mg·h·L^-1，AUC0-∞为（1．23±0．35）mg·h·L^-1。结论该方法适用于格拉司琼贴剂在巴马小型猪体内药物动力学行为的研究，贴剂可延长格拉司琼作用时间。

Objective To develop a chromatography-tandem mass spectrometry （LC-MS/MS） method for the determination of granisetron in pig plasma, and study the pharmaceutical characteristic of granisetron in pig plasma. Methods The chromatographic separation was accomplished on a Diamonsil C18 analytical column （150 mm×4. 6 mm,5μm） at a flow rate of 0. 5 mL·min^-1 and the mobile phase was consisted of methanol-5 mmol· L^ - 1 ammonium acetate-formic acid （ V： V： V = 45： 55： 0. 1 ）. Liquid-liquid extraction was used for the sample pretreatment with a plasma volume of 200 μL. The plasma concentration was detected by multiple reaction monitoring after the pig was given granisetron patch. Results The method was linear over the range of 0. 020- 20. 0 μg· L^- 1 and the lower limit of quantification （LLOQ） was 20 ng· L^ - 1. The intra- day and inter-day precision was within the limits of 13.4% in terms of relative standard deviation （RSD%）. The average recoveries were between 63.8%-66. 7%. The matrix effects were between 101.4%- 105.8%. The Main pharmacokinetic parameters were as follows ： t1/2 （ 18.0 ± 1.4 ） h ,ρmax （ 15.3± 4.0） μg·L^-1,MRT （91.9 ±4.9） h,Kc（0.04 ±0.00） h^-1,AUC0-216（1.22 ±0.35） mg·h·L^-1,AUC0-∞ （ 1.23 ±0. 35 ） mg· h· L^- 1 , respectively. Conclusions This simple and sensitive LC-MS/MS method has been successfully applied to pharmacokinetic studies of granisetron in pigs after a patch.