用高效液相色谱-质谱-质谱法测定小鼠血浆和大脑中的石杉碱甲及药动学  

Determination and study the pharmcokinetic of hupernize A in mouse plasma and brain by HPLC/MS/MS

作　　者：刘骁[1] 李端[1] 王小林 陈春麟 

机构地区：[1]复旦大学药学院药理学教研室,上海200032 [2]上海美迪西生物医药有限公司,上海201203

出　　处：《复旦学报（医学版）》2006年第2期247-250,共4页Fudan University Journal of Medical Sciences

摘　　要：目的建立小鼠血浆和大脑中石杉碱甲的液质联用测定法，测定灌胃给药后小鼠血浆和大脑中石杉碱甲浓度的动态变化以及药动学参数。方法小鼠血浆及大脑样品经液液萃取，用高效液相色谱-质谱-质谱（LC-MS-MS）联用法检测。色谱柱：PHENOMENEX C18柱，100mm×2mm，5micron；流动相为甲醇：水（1％甲酸溶液）=60：40；用于定量的离子为m／z243→154（石杉碱甲）和m／z278→203（内标：克仑特罗）。结果所建立的血浆的方法线性范围为1～1000ng／mL，脑的方法线性范围为1～1000ng／mL，它们最低检测限为1ng／mL，（r=0．9990），RSD〈7％，可以检测小鼠灌胃后石杉碱甲在血和脑中的浓度；血浆石杉碱甲浓度-时间变化可用一室模型拟合，主要药动学参数分别为Tmax=6min，Cmax=281ng／mL，AUC（0→tn）=35266ng·L^-1·min^-1，AUC（0→∞）=38056ng·L-^1·min^-1，t1／2=99min；在大脑中的药物浓度-时间变化用非房室模型拟合，主要药动学参数分别为Tmax=60min，Cmax=224ng／mL，AUC（0→tn）=44344ng·L^-1·min^-1，AUC（0→∞）=58571ng·L^-1·min^-1。结论该方法灵敏度高，简便快速，可用于药代动力学及相关研究；石杉碱甲在小鼠大脑中的消除速度比血浆缓慢。Purpose An HPLC/MS/MS method was developed to measure the concentration of hupernize A in mouse plasma and brain. The method was used to preliminarily study pharmacokinetics of hupernize A in mouse plasma and brain. Methods Samples were extracted liqued-liqued extraction and separated on PHENOMENEX C18 column. The mobile phase is methanol： water = 60：40. Quantitation was based on multiple reaction monitoring（MRM） using the precursor→production combination of m/z 2 43→154 and m/z 278→203 for Hupernize A and Clenbuterol respectively. Results 1. The method has a linear range from 1 - 1 000 ng·mL^-1 （r = 0. 999 0） and minimal detectable concentration at 1 ng/ml. The intra-day and inter-day RSD were less than 7%. The method can be applied to determine plasma concentrations and brain concentrations. 2. The plasma concentrations vs time curve was fitted as an open one-compartment model. The pharmacokinetic parameters of mouse plasma were 6min for Tmax,281 ng·mL^-1 for Cmax,35 266 ng·L^-1·min^-1 for AUC（0→tn） ,AUC（0→∞） =38 056 ng·L^-1·min^-1 and t1/2 = 99 min; The brain concentrations vs time curve was fitted as an noncompartment model. The pharmacokinetic parameters of mouse brain were 60min for Tmax, 224 ng·mL^-1 for C 44344 ng·L^-1·min^-1 for AUC（0→tn） and AUC（0→∞） = 58 571 ng·L^-1·min^-1. Conclusions The method is sensitive, simple and rapid. It can be applied to study the pharmacokinetics of hupernize A.Hupernize A was eliminated more slowly in mouse brain than in mouse plasma.

关 键 词：石杉碱甲 高效液相-质谱-质谱联用法 药动学 

分 类 号：R969.1[医药卫生—药理学]