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高效液相-质谱串联方法快速测定大鼠血浆中的槲皮素-3'-O-葡萄糖苷含量

黄慧莲, 刘科兰, 邵峰, 任刚, 叶耀辉, 张寿文, 刘荣华*   

  1. 江西中医学院 现代中药制剂教育部重点实验室, 江西 南昌 330004
  • 收稿日期:2011-12-21 修回日期:2012-03-12 出版日期:2012-05-15 发布日期:2012-05-15
  • 通讯作者: 刘荣华*

Rapid determination of quercetin-3'-O-glucoside in rat plasma by high
performance liquid chromatography-tandem mass spectrometry

Hui-Lian Huang, Ke-Lan Liu, Feng Shao, Gang Ren, Yao-Hui Ye, Shou-Wen Zhang, Rong-Hua Liu*   

  1. Key Laboratory of Modern Preparation of Traditional Chinese Medicine, Jiangxi University of Traditional Chinese Medicine, Ministry of Education, Nanchang 330004, China
  • Received:2011-12-21 Revised:2012-03-12 Online:2012-05-15 Published:2012-05-15
  • Contact: Rong-Hua Liu*

摘要:

建立大鼠血浆中槲皮素-3'-O-葡萄糖苷的高效液相-串联质谱 (HPLC-MS/MS) 检测法。生物样品中槲皮素-3'-O-葡萄糖苷和卡马西平 (内标) 采用液-液萃取的方法提取, 乙酸乙酯作为提取溶剂。HPLC-MS/MS方法采用多离子反应模式 (MRM) 进行检测, 被测物质和内标均在3.5 min内出峰。槲皮素-3'-O-葡萄糖苷在血浆中的最低定量限是10.625 ng/mL, 最低检测限是4.25 ng/mL, 平均回收率大于66.80%, 日内、日间精密度和准确度的RSD值均小于10.44%。尾静脉注射10 mg/kg槲皮素-3'-O-葡萄糖苷后, 其 t1/2 AUC分别是 (0.02±0.01) h 和 (1.22±0.28)×104 μg/L·h。本法准确、稳定、灵敏度高, 适用于槲皮素-3'-O-葡萄糖苷的大鼠药代动力学研究。

关键词: 高效液相-质谱串联法, 槲皮素-3'-O-葡萄糖苷, 血药浓度, 药代动力学

Abstract: A method for the quantification of quercetin-3'-O-glucoside in rat plasma by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated. Along with internal standard (carbamazepine), quercetin-3'-O-glucoside was extracted from plasma samples by simple liquid-liquid extraction with ethyl acetate. The mass spectrometry detection was set in multiple reaction monitoring (MRM) mode via positive electrospray ionization (ESI). The chromatographic run time was 3.5 min per sample. The calibration curves were linear (r2 = 0.9992) with a lower limit of quantification (LLOQ) of 10.625 ng/mL, and the limit of detection (LOD) was 4.25 ng/mL. The intra- and inter- day precision and accuracy, in terms of relative standard deviation (RSD), were all lower than 10.44%. The recovery rate of the analyte and internal standard were higher than 66.80%. After intravenous administration of 10 mg/kg quercetin-3'-O-glucoside, the t1/2 and AUC were (0.02±0.01) h and (1.22±0.28)×104 μg/L·h. The method is accurate, stable and sensitive, which is suitable for the pharmacokinetic study of quercetin-3'-O-glucoside in rats.

Key words: HPLC-MS/MS, Quercetin-3'-O-glucoside, Plasma concentration, Pharmacokinetics

中图分类号: 

Supporting:

Foundation items: National Natural Science Foundation of China (Grant No. 30960494) and Jiangxi Province NSFC (Grant No. 2009 GZY0099).
*Corresponding author. Tel.: 86-791-87118658