液质联用法测定犬血浆中阿普唑仑及其代谢物α-羟基阿普唑仑的浓度  

Determination of alprazolam and its metaboliteα-hydroxy-alprazolam in plasma of beagle dogs by LC-MS

作　　者：毛叶萌[1,2] 张奇志[1] 武炳贤[1] 沈烨虹[1] 陈恩[1] 冯亮[1] 蒋新国[1] 

机构地区：[1]复旦大学药学院药剂学教研室,上海200032 [2]上海市精神卫生中心,上海200030

出　　处：《中国新药与临床杂志》2008年第10期746-750,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：上海市卫生局资助项目(2006065)

摘　　要：目的建立测定Beagle犬血浆中阿普唑仑及其代谢物α-羟基阿普唑仑浓度的液相色谱-质谱联用(LC-MS)法。方法血浆样品采用1 mol·L^(-1)硼酸盐缓冲液(pH 9.0)碱化、乙酸乙酯-正庚烷(85:15,V:V)萃取后LC-MS测定。色谱柱:Zorbax SB-C_(18)柱(150 mm×3 mm,3.5μm);流动相:乙腈-0.01 mol·L^(-1)乙酸胺缓冲液(含1%甲酸)(45:55,V:V);流速:0.3 mL·min^(-1);柱温:40℃。采用电喷雾正离子模式离子化,用于定量分析的离子分别为m/z 309.2(阿普唑仑)、m/z 325.2(α-羟基阿普唑仑)和m/z 343.2(三唑仑,内标)。结果阿普唑仑和α-羟基阿普唑仑的线性范围分别为0.5～50μg·L^(-1)和0.5～32μg·L^(-1),两者定量下限均为0.5μg·L^(-1),提取回收率均>80%,方法回收率为97.3%～102.5%,批内RSD≤10.4%,批间RSD≤12.2%。结论本方法灵敏、准确、重现性好,适用于阿普唑仑犬体内药动学研究。AIM To develop a LC-MS method for the determination of alprazolam and its metabolite α- hydroxy-alprazolam in plasma of beagle dogs. METHODS The plasma samples were alkalinized with 1 mol·L^-1 borate buffer （pH = 9.0） and extracted with acetoacetate-n-heptane （85 ： 15, V ： V）. Chromatographic separation was conducted at 40℃ with a Zorbax SB C18 （150 mm × 3 mm, 3.5 um） analytical column. The mobile phase consisted of acetonitrile -0.01 mol·L^-1 ammonium acetate containing 1% formic acid （45 ： 55, V ： V） at a flow rate of 0.3 mL·min^-1. Ion mass spectral （m/z） of 309.2, 325.2, and 343.2 were selected to quantify alprazolam, α-hydroxy-alprazolam and triazolam （internal standard）, respectively. RESULTS The calibration curves were linear over the range of 0.5 - 50ug·L^-1 and 0.5 - 32 ug·L^-1 for alprazolam and α- hydroxy-alprazolam, respectively. The lower limits of quantification were 0.5ug·L^-1, and the extraction recoveries of alprazolam and its metabolite from plasma were more than 80 %. Method recovery was 97 - 102.5 % and within-and between-batch variations were less than 10.4 % and 12.2 %, respectively. CONCLUSION The established method is sensitive, accurate and reproducible, and suitable for the pharmacokinetic study of alprazolam tablets.

关 键 词：阿普唑仑 α-羟基阿普唑仑 血浆 犬 色谱法 高压液相 光谱法 质量 电喷雾电离 

分 类 号：R969.1[医药卫生—药理学]