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LC-ESI-MS/MS法测定人血浆中卢帕他定的浓度

温预关*, 喻凌寒, 彭建玲, 廖日房, 马崔   

  1. 1. 广州市脑科医院国家药品临床研究基地, 广东 广州510370;
    2. 广东省化学危害应急检测技术重点实验室(中国广州分析测试中心), 广东 广州 510070;
    3.中山大学附属第二医院临床药学部, 广东 广州 510120
  • 收稿日期:2007-01-15 修回日期:2007-05-10 出版日期:2007-06-15 发布日期:2007-06-15
  • 通讯作者: 温预关*

Sensitive liquid chromatography electrospray ionization ion-trap mass spectrometry for the determination of rupatadine in human plasma

Yu-Guan Wen*, Ling-Han Yu, Jian-Ling Peng, Ri-Fang Liao, Cui Ma   

  1. 1.Department of Clinical Pharmacology, Guangzhou Brain Hospital, Guangzhou 510370, China;
    2.Guangdong Key Laboratory of Chemical Emergency Test, China National Analytical Center, Guangzhou 510070, China;
    3.Department of pharmacy, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China

  • Received:2007-01-15 Revised:2007-05-10 Online:2007-06-15 Published:2007-06-15
  • Contact: Yu-Guan Wen*

摘要: 目的 建立测定人血浆中卢帕他定浓度的LC-ESI-MS/MS方法。方法 血浆样品加入内标, 碱化后用二氯甲烷: 乙酸乙酯(20:80) 提取, 37 ºC真空干燥箱中干燥至干,残渣用200 μL流动相溶解后进样。色谱条件为:色谱柱为Agilent EclipseXDB-C18 (4.6 mm × 150 mm,5 μm); 流动相为乙腈(1%甲酸): 20 mmol·L–1 醋酸铵(76:24,V/V), 流速为0.6 mL·min–1。质谱条件:采用美国安捷仑1100高效液相色谱系统和离子阱(Agilent MSD Trap XCT)检测仪, 质谱条件为电喷雾离子源,检测方式为正离子电离、多离子反应监测(MRM), 用于定量分析的离子为卢帕他定m/z 416→309, 内标氯雷他定m/z 383→337结果 该方法应用于检测20名健康志愿者服药后的血浆样品。线性范围为0.05 ~14 ng·mL–1 (r = 0.998), 日内和日间精密度均低于15%,方法回收率为85.1% ~ 114.0%。最低检测限为0.05 ng·mL–1 (n = 5,RSD = 9.22%)结论 该方法灵敏、准确、快速,可用于该药药代动力学和生物等效性研究。

关键词: 卢帕他定, 卢帕他定, 卢帕他定, HPLC-ESI-MS/MS, HPLC-ESI-MS/MS, HPLC-ESI-MS/MS, 血浆, 血浆, 血浆, 药代动力学, 药代动力学, 药代动力学

Abstract: Aim To develop and validate a sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry (LC-ESI-MS/MS) method for the identification and concentration of rupatadine in human plasma. Methods After the addition of the internal standard (IS, loratadine) and 0.01 mol·L–1 sodium hydroxide solution, plasma samples were extracted with methylene chloride: ethyl acetate mixture (20:80, V/V). The organic layer was evaporated under vacuum drying at 37 ºC. The residue was reconstituted with 200 μL mobile phase. Chromatography was performed on an Agilent Eclipse XDB-C18 (4.6 mm × 150 mm, 5 μm) column with a mobile phase consisting of acetonitrile (1% formic acid) –20 mmol·L–1 ammonium acetate (76:24, V/V) at a flow-rate of 0.6 mL·min–1. Detection was performed on Agilent MSD Trap XCT ion-trap mass spectrometry connected to a Agilent 1100 high performance liquid chromatography (HPLC) by selected reaction monitoring (SRM) mode via electrospray ionization (ESI) source. Rupatadine (MRM m/z 416309) and loratadine (MRM m/z 383337) were detected by Agilent MSD Trap XCT ion-trap mass analyser. Results The method was proved to be sensitive and specific by testing 20 different plasma batches. Linearity was established for the range of concentrations 0.05 – 14.0 ng·mL–1 with a coefficient of determination (r) of 0.998. The intra-and inter-day precision (RSD %) were lower than 15% and accuracy ranged from 85.1% to 114.0%. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.05 ng·mL–1 with a precision of 9.22% (n = 5). Conclusion The proposed method is sensitive and reproducible enough to be used in pharmacokinetic, bioavailability or bioequivalence studies of rupatadine.

Key words: Rupatadine, Rupatadine, HPLC-ESI-MS/MS, HPLC-ESI-MS/MS, Human plasma, Human plasma, Pharmacokinetics, Pharmacokinetics

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Supporting: *Corresponding author. Tel.: 86-20-81891624; fax: 86-20-81891391