色谱

色谱 ›› 2014, Vol. 32 ›› Issue (7): 779-783.DOI: 10.3724/SP.J.1123.2014.03030

• 技术与应用 • 上一篇    下一篇

同位素稀释-气相色谱-质谱法检测大鼠血浆中的内源性胍丁胺

丘忠丽1,2, 林缨1, 熊志立2, 谢剑炜1   

  1. 1. 抗毒药物与毒理学国家重点实验室, 军事医学科学院毒物药物研究所, 北京 100850;
    2. 沈阳药科大学药学院, 辽宁 沈阳 110016
  • 收稿日期:2014-03-19 修回日期:2014-05-07 出版日期:2014-07-08 发布日期:2014-06-25
  • 通讯作者: 谢剑炜，E-mail：xiejw@bmi.ac.cn；Tel:(024)23986289；熊志立，E-mail：beary200@126.com.

Determination of endogenous agmatine in rat plasma by isotope dilution-gas chromatography-mass spectrometry

QIU Zhongli1,2, LIN Ying1, XIONG Zhili2, XIE Jianwei1   

  1. 1. State Key Laboratory of Toxicology and Medical Countermeasures, and Laboratory of Toxicant Analysis, Institute of Pharmacology and Toxicology, Academy of Military Medical Science, Beijing 100850, China;
    2. College of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
  • Received:2014-03-19 Revised:2014-05-07 Online:2014-07-08 Published:2014-06-25

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321

被引次数

4

摘要：

建立了大鼠血浆中内源性胍丁胺的同位素稀释-气相色谱-负化学电离质谱定量分析方法。大鼠血浆样品经蛋白沉淀并蒸干后，用六氟乙酰丙酮衍生化，采用Florisil固相萃取柱净化，以稳定同位素标记的d8-胍丁胺为内标，在气相色谱-质谱仪上采用负化学电离方式电离，选择离子模式检测。胍丁胺标准溶液的检出限为0.0057 ng/mL。血浆中添加的胍丁胺在1.14~57.0 ng/mL范围内呈良好的线性关系（r=0.997），方法回收率介于92.3%~109.8%之间，日内和日间精密度均小于15%。大鼠血浆中胍丁胺平均含量水平为（22±9）ng/mL，雌、雄大鼠血浆中的胍丁胺水平未见显著性差异（p>0.05）。该方法特异性好、灵敏度高，为生物体内胍丁胺的生理功能研究提供了高灵敏的分析方法。

关键词: 大鼠, 内源性胍丁胺, 气相色谱-负离子化学电离质谱, 同位素稀释, 血浆

Abstract:

A method for the determination of endogenous agmatine in rat plasma was developed by isotope dilution-gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS). The plasma samples were analyzed after protein precipitation, evaporation, derivatization by hexafluoroacetone (HFAA), and clean-up on a Florisil SPE column. The GC-MS analysis utilized stable isotope d8-agmatine as internal standard. The samples after treatment were tested by negative chemical ionization with selected ion monitoring (SIM) which was set at m/z 492 (molecular ion of agmatine) and m/z 500 (molecular ion of internal standard). The limit of detection (LOD) of agmatine standard solution was 0.0057 ng/mL. The calibration curve of the agmatine spiked in rat plasma showed a good linear relationship at the range of 1.14-57.0 ng/mL (r=0.997). The recoveries of agmatine spiked in rat plasma ranged from 92.3% to 109.8%. Inter-day and intra-day precisions were less than 15%. The average concentration level of agmatine in rat plasma was (22±9) ng/mL, and there was no significant difference between male and female SD rats (p>0.05). The method is high sensitive and specific, and can be used for the determination of endogenous agmatine in plasma. It provides a strong support for the subsequent research of agmatine.

Key words: endogenous agmatine, gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), isotope-dilution, plasma, rat

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