色谱

色谱 ›› 2011, Vol. 29 ›› Issue (12): 1205-1209.DOI: 10.3724/SP.J.1123.2011.01205

• 研究论文 • 上一篇    下一篇

柱切换技术-高效液相色谱法快速检测大鼠血浆中的普萘洛尔对映体

武晓玉1, 王荣1,2*, 谢华1, 王剑锋1, 贾正平1,2*, 张强1, 王先华1   

  1. 1. 兰州军区兰州总医院临床药理基地, 甘肃 兰州 730050; 2. 兰州大学药学院, 甘肃 兰州 730000
  • 收稿日期:2011-08-01 修回日期:2011-09-02 出版日期:2011-12-28 发布日期:2012-01-15
  • 通讯作者: 王荣,教授,主要研究方向为药物体内分析. E-mail: wangrong-69@163.com.贾正平,教授,博士生导师,主要研究方向为手性药物分析及分子药理学. Tel: (0931)8994652.
  • 基金资助:

    国家科技部重大项目(No. 2008ZXJ09014-010)

Rapid determination of propranolol enantiomers in rat plasma by column-switching-high performance liquid chromatography

WU Xiaoyu1, WANG Rong1,2*, XIE Hua1, WANG Jianfeng1, JIA Zhengping1,2*, ZHANG Qiang1, WANG Xianhua1   

  1. 1. Base for Clinical Pharmacology, Lanzhou General Hospital of Lanzhou Military Region, Lanzhou 730050, China; 2. Pharmacy college, Lanzhou University, Lanzhou 730000, China
  • Received:2011-08-01 Revised:2011-09-02 Online:2011-12-28 Published:2012-01-15

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840

被引次数

27

摘要： 建立了快速检测大鼠血浆中普萘洛尔对映体浓度的柱切换-高效液相色谱法。将自制限进填料柱作为预处理柱,通过直接进样方式,使普萘洛尔对映体在预处理柱上保留,同时除去血浆中的蛋白质等大分子;再通过柱切换技术,使普萘洛尔对映体在键合型纤维素-三(3,5-二甲基苯基氨基甲酸酯)(Chiralcel OD-RH)分析柱上得到手性拆分。通过条件优化,确定切换前预处理流动相为硼酸盐缓冲液(pH 8.5)-甲醇(95:5, v/v),流速为1.0 mL/min;切换后分析流动相为异丙醇-乙醇-0.2 mmol/L硼酸盐缓冲液(pH 8.5)(30:30:40, v/v/v),流速为0.8 mL/min;切换时间为3 min;柱温为25 ℃;检测波长为293 nm。普萘洛尔两对映体在25～500 mg/L的质量浓度范围内具有良好的线性关系(r=0.9995), 3个加标水平(50、100、250 mg/L)的平均回收率为97.89%～101.56%,日内和日间精密度均小于5%。该方法简便、快速、灵敏、准确,适于血浆样本中手性药物的药代动力学研究。

关键词: 大鼠血浆, 高效液相色谱, 普萘洛尔对映体, 限进填料, 药代动力学, 柱切换

Abstract: A high performance liquid chromatographic (HPLC) method with column-switching was developed and validated for rapid determination of two propranolol enantiomers in rat plasma. The column of restricted-access media was used as a pre-treatment column and a Chiralcel OD-RH was used as analytical column. The plasma samples were injected directly into the pre-treatment column to remove plasma protein and endogenous constituents as well as to retent the propranolol enantiomers in the column using the mobile phase of borate buffer (pH 8.5)-methanol (95:5, v/v) at the flow rate of 1.0 mL/min. Then the propranolol enantiomers were transferred to the Chiralcel OD-RH column using the mobile phase of isopropanol-ethanol-0.2 mmol/L borate buffer (pH 8.5) (30:30:40, v/v/v) at a flow rate of 0.8 mL/min by column-switching technology. The column-switching time was 3.0 min, the used wavelength was 293 nm and the column temperature was set at 25 ℃. The calibration curve showed excellent linear relationship (r=0.9995) in the concentration range from 25 mg/L to 500 mg/L for propranolol enantiomers in plasma. The intra-day and inter-day assay precisions and accuracies were well and the relative standard deviations (RSDs) were less than 5%. The average recoveries (n=6) of the two enantiomers at 3 spiked levels were from 97.89% to 101.56%. All the values of the method validation were within the generally accepted criteria for biological sample analysis. The results show that the method is convenient, quick, sensitive and accurate. The method was successfully applied in the determination of propranolol enantiomers in rat blood pharmacokinetics study.

Key words: high performance liquid chromatography (HPLC), pharmacokinetics, propranolol enantiomers, rat plasma, restricted-access media, column-switching