高等学校化学学报

高等学校化学学报

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7-甲氧基香豆素-3-羧酸-N-琥珀酰亚胺酯用于神经肽的标记与分析

李军1, 刘祥军1, 赵睿1, 熊少祥1, 马会民1, 吕强2, 许莉2   

    1. 中国科学院化学研究所, 北京 100080;
    2. 北京微量化学研究所, 北京 100091
  • 收稿日期:2006-02-28 修回日期:1900-01-01 出版日期:2006-12-10 发布日期:2006-12-10
  • 通讯作者: 马会民

Labeling Analysis of Neuropeptides with 7-Methoxycoumarin-3-carboxylic Acid N-Succinimidyl Ester

LI Jun1, LIU Xiang-Jun1, ZHAO Rui1, XIONG Shao-Xiang1, MA Hui-Min1, L<SPAN lang=EN-US style=   

    1. Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China;
    2. Beijing Institute of Microchemistry, Beijing 100091, China
  • Received:2006-02-28 Revised:1900-01-01 Online:2006-12-10 Published:2006-12-10
  • Contact: MA Hui-Min

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摘要： 选用7-甲氧基香豆素-3-羧基-N-琥珀酰亚胺酯(MCSE)作为衍生试剂, 并借助高效液相色谱和质谱等仪器对甲硫脑啡肽、亮脑啡肽和神经紧张素等3种神经肽进行了标记与分析.

关键词: 神经肽, 标记分析, 7-甲氧基香豆素-3-羧基-N-琥珀酰亚胺酯

Abstract: Labeling analysis of three neuropeptides(Met-enkephalin, Leu-enkephalin, and neurotensin) with 7-methoxycoumarin-3-carboxylic acid N-succinimidyl ester(MCSE) was investigated. The experimental conditions for labeling reaction and HPLC analysis of the neuropeptide derivatives were optimized. It was found that the labeling reatction could be achieved at 37 ℃ for one hour in H3BO3-NaCl-Na2B4O7 buffer(pH=83) with a labeling ratio of about 80%. The derivative mixture from the three peptides can be separated in a 40 min linear gradient from 60% to 20% of solvent A(100 mmol/L sodium acetate, pH=50) and from 40% to 80% of solvent B[60%(volume fraction) acetonitrile in water]. The resulting labeled products were also confirmed by MALDI-TOF-MS analysis. Compared with the native neuropeptides, the resulting labeled products have a longer absorption and fluorescence wavelengths, which is beneficial to the effective elimination of background fluorescence interference from biological matrix.

Key words: Neuropeptides, Labeling analysis, 7-Methoxycoumarin-3-carboxylic acid N-succimidyl ester

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