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

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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

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