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固相萃取前处理HPLC-MS法测定口服给药后家兔血浆中的灯盏乙素

李妮, 黄建明, 翁伟宇, 黄照昌, 蔡佳, 郁韵秋*   

  1. 1.复旦大学药学院, 上海 200032;
    2.昆明制药集团股份有限公司, 云南 昆明 650100

  • 收稿日期:2004-11-22 修回日期:2005-02-10 出版日期:2005-03-15 发布日期:2005-03-15

Determination of Scutellarin in Rabbit Plasma after Oral Administration by HPLC-MS with Solid-Phase Extraction

LI Ni, HUANG Jian-ming, WENG Wei-yu, HUANG Zhao-chang, CAI Jia, YU Yun-qiu*   

  1. 1. School of Pharmacy, Fudan University, Shanghai 200032, China;
    2. Kunming Ph armaceutical Corporation, Kunming 650100, China
  • Received:2004-11-22 Revised:2005-02-10 Online:2005-03-15 Published:2005-03-15

摘要: 目的 建立灵敏、专属的HPLC-ESI-MS测定法, 用于测定家兔口服灯盏花素缓释片(2 60 mg/)后的血药浓度。方法 该定量方法采用芦丁作内标, 用固相萃取(SPE)进行样品前处理, 以甲醇10 mmol·L-1醋酸铵水溶液(pH 8.0)梯度洗脱, 流动相流速0.4 mL·min-1, 柱温35 ºC, 经过Zorbax Extend-C18 (150 mm×2.1 mm ID, 5 μm)色谱柱分离, 电喷雾质谱检测器在负离子模式下检测灯盏乙素和芦丁。结果 2200 ng·mL-1范围内灯盏乙素与内标峰面积与浓度线性关系良好, 且准确度、精密度满足生物样品分析的要求。结论 本实验建立的兔血浆中灯盏乙素的HPLC-MS分析方法灵敏度高, 可用于口服给药后灯盏乙素的血药浓度测定.

关键词: 灯盏乙素, 灯盏乙素, 灯盏乙素, 液质联用, 液质联用, 液质联用, 固相萃取, 固相萃取, 固相萃取, 灯盏花素, 灯盏花素, 灯盏花素

Abstract: Aim To establish a sensitive and specific liquid chromatography-mass s pectrometry method for determination of scutellarin in rabbit plasma after oral administration. Methods For the quantitative analysis, rutin was used as an internal standard and solid-phase extraction (SPE) was performe d by using a Phenomenex C8 cartridge. HPLC was carried out using a Zorbax Exte nd-C18 column (150 mm×2.1 mm ID, 5 μm) with a guard cartridge (Phenomenex). Gradient elution was selected with the mobile phase of methanol 10 mmol·L-1 ammonium acetate solution (pH adjusted to 8.0 with ammonia soluti on). The flow rate of mobile phase was 0.4 mL·min-1 and the column temperatur e was 35 ºC. Both scutellarin a nd the internal standard rutin in rabbit plasma extracts were detected by mass s pectrometry using an ESI interface in the negative ion mode. Results The linear range was from 2 to 200 ng·mL-1, with acceptable accuracy and precision (RSD). Conclusion A sensitive, reliable and accurate method for the quantitati on of scutellarin in rabbit plasma has been established.

Key words: scutellarin, scutellarin, LC-MS, LC-MS, SPE, SPE, breviscapine, breviscapine

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