色谱

色谱 ›› 2012, Vol. 30 ›› Issue (07): 705-710.DOI: 10.3724/SP.J.1123.2012.03001

• 研究论文 • 上一篇    下一篇

酸化沉淀蛋白-超快速液相色谱-串联质谱法测定肝损伤大鼠血浆中的头孢呋辛

赵龙山1, 李清1, 杨威2, 何博赛1, 魏斌斌1, 刘然1, 刘晶晶1, 陈晓辉1, 毕开顺1*   

  1. 1. 沈阳药科大学药学院, 辽宁 沈阳 110016; 2. 广州医药研究总院药物非临床评价研究中心, 广东 广州 510663
  • 收稿日期:2012-03-01 修回日期:2012-04-11 出版日期:2012-07-28 发布日期:2012-07-19
  • 通讯作者: 毕开顺,博士生导师,主要从事药物分析研究. Tel: (024)23986012, E-mail: bikaishun@yahoo.com.
  • 基金资助:

    国家“重大新药创制”科技重大专项(2009ZX09301-012).

Determination of cefuroxime in liver-injured rat plasma by ultra fast liquid chromatography-tandem mass spectrometry via acidified protein precipitation

ZHAO Longshan1, LI Qing1, YANG Wei2, HE Bosai1, WEI Binbin1, LIU Ran1, LIU Jingjing1, CHEN Xiaohui1, BI Kaishun1*   

  1. 1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China; 2. Center for Drug Non-clinical Evaluation and Research of Guangzhou General Pharmaceutical Research Institute, Guangzhou 510663, China
  • Received:2012-03-01 Revised:2012-04-11 Online:2012-07-28 Published:2012-07-19

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摘要： 为了研究二代头孢类新药头孢呋辛赖氨酸在肝损伤大鼠体内的药代动力学过程,建立了采用超快速液相色谱-串联质谱(UFLC-MS/MS)快速测定肝损伤模型大鼠血浆中头孢呋辛含量的方法。血浆样品在酸性条件下用乙腈沉淀蛋白,采用Shim-pack XR-ODS色谱柱(75 mm×3.0 mm, 2.2 μm)为分析柱、乙腈-0.1%甲酸水溶液(40:60, v/v)为流动相、流速为400 μL/min进行色谱分离,采用电喷雾负离子(ESI~)模式电离、多反应监测(MRM)模式进行质谱检测,用于定量分析的离子对分别为m/z 423.2→206.8 (头孢呋辛)和m/z 454.1→238.4 (内标头孢噻肟)。结果表明,大鼠血浆中头孢呋辛的质量浓度在0.01～1 mg/L和1～400 mg/L范围内线性关系良好(r＞0.99),定量限为0.01 mg/L,日内和日间精密度(以相对标准偏差(RSD)计)均小于11.5%,准确度(RE)为~7.1%～2.2%,平均萃取回收率大于83.5%,样品运行时间仅为3.0 min,能够满足生物样品的测定需求。该法简便、快速,已用于肝损伤大鼠静脉注射头孢呋辛赖氨酸的药代动力学预实验研究。

关键词: 超快速液相色谱-串联质谱法, 酸化沉淀蛋白, 头孢呋辛, 血药浓度

Abstract: In order to investigate the pharmacokinetic profiles of cefuroxime lysine, a new second generation cephalosporins, in liver-injured rat model, an ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method for the determination of cefuroxime in liver-injured rat plasma was developed and validated. The plasma sample was pretreated by protein precipitation with acidified acetonitrile. The analytes were separated on a Shim-pack XR-ODS column (75 mm×3.0 mm, 2.2 μm) with acetonitrile-0.1% formic acid aqueous solution (40:60, v/v) as the mobile phase at a flow rate of 400 μL/min. The mass spectrometer was operated in multiple reaction monitoring (MRM) mode with a negative electrospray ionization (ESI) interface. The precursor to product ion transitions of m/z 423.2→206.8 and m/z 454.1→238.4 were selected to determine cefuroxime and cefotaxime (internal standard, IS), respectively. The linearities ranged from 0.01 to 1 mg/L and 1 to 400 mg/L (r＞0.99), and the limit of quantification of cefuroxime was 0.01 mg/L. The relative standard deviations (RSDs) of intra- and inter-day precisions were both less than 11.5%, and the accuracy (relative error) was between ~7.1% and 2.2%. The mean extraction recovery was more than 83.5%. The total run time was 3.0 min per sample. The method is simple and fast for the preliminary pharmacokinetic study of cefuroxime lysine in liver-injured rats.

Key words: acidified protein precipitation, cefuroxime, plasma drug concentration, ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS)

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