色谱

色谱

• 研究论文 • 上一篇    下一篇

高效液相色谱法测定大鼠给药盐酸氟西汀后脑渗析液中的5-羟色胺

郭云珍,于莉,马郑,郭兴杰

  

  1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
  • 收稿日期:2006-07-22 修回日期:2006-11-21 出版日期:2007-03-30 发布日期:1985-12-25
  • 通讯作者: 郭兴杰

Determination of Serotonin in Rat Brain Microdialysate After i.p. Administration of Fluoxetine Hydrochloride byHigh Performance Liquid Chromatography

GUO Yunzhen, YU Li, MA Zheng, GUO Xingjie   

  1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
  • Received:2006-07-22 Revised:2006-11-21 Online:2007-03-30 Published:1985-12-25

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摘要： 建立了高效液相色谱柱前衍生化法测定大鼠腹腔注射(i.p.)给药盐酸氟西汀前后脑渗析液中5-羟色胺(5-HT)的浓度变化情况。在大鼠给药盐酸氟西汀前和给药后不同时间点取其脑渗析液,加入衍生化试剂反应后,以乙腈-20 mmol/L醋酸盐缓冲液(pH 5.0)(体积比为45∶55)（含20 mmol/L辛烷磺酸钠）作为流动相,在Hypersil C18色谱柱 (250 mm×2.0 mm,5 μm)上进行分离,荧光检测波长为λex=330 nm，λem=455 nm。5-HT在浓度为0.25～5.0 nmol/L 范围内线性关系良好(r=0.9991);5-HT的最低定量限为0.25 nmol/L。本法准确可靠、简便,适用于生物样品中5-HT的浓度测定。

关键词: 5-羟色胺, 高效液相色谱, 盐酸氟西汀 , 荧光检测, 柱前衍生化

Abstract: The concentration of serotonin (5-HT) in rat brain microdialysate before and after i.p. administration of fluoxetine hydrochloride was determined through precolumn derivatization by reversed-phase high performance liquid chromatography (RP-HPLC). A baseline separation of serotonin was achieved on a Hypersil C18 column (250 mm×2.0 mm, 5 μm) with acetonitrile-20 mmol/L sodium acetic solution (pH 5.0) (45∶55, v/v) containing 20 mmol/L of sodium octanesulfonate as the mobile phase. Fluorescence detection with the excitation wavelength at 330 nm and emission wavelength at 455 nm was used for serotonin. There was good linear relationship between the peak area and concentration in the range of 0.25-5.0 nmol/L(r=0.9991). The limit of quantitation was 0.25 nmol/L. The method is simple, accurate and can be applied to the determination of serotonin in biological specimen.

Key words: fluorescence detection, fluoxetine hydrochloride , high performance liquid chromatography (HPLC), serotonin, precolumn derivatization