寡糖衍生化及基质辅助激光解吸电离飞行时间质谱分析方法研究

Derivatization of Oligosaccharides and Analysis by Matrix-assisted Laser Desorption/Ionization-Time of Flight-Mass Spectrometry

为提高中性寡糖在基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)中的检测灵敏度,建立了以1-(4-氰基苯基)-4-哌啶碳酰肼(CPH)为衍生化试剂对寡糖的标记方法。寡糖的还原端与CPH的酰肼基团反应生成腙,使得寡糖被CPH标记,衍生物以MALDI-TOF质谱进行分析。结果表明:在反应温度95℃,醋酸浓度为0.125%(V/V),CPH过量100倍的条件下,衍生产率可达最大,并且CPH衍生可使中性寡糖在MALDI-TOF质谱中的检测灵敏度提高10倍。本方法简便快速,灵敏度高,适合微量寡糖链的质谱分析。

A derivatization method for the determination of oligosaccharides with high sensitivity by matrixassisted laser desorption/ionization-time of flight-mass spectrometry（ MALDI-TOF-MS） based on a labeling regent 1-（ 4-Cyanophenyl）-4-piperidinecarbohydrazide（ CPH） was proposed. The hydrazones were generated by the reaction of the reducing ends of oligosaccharides and the hydrazide of CPH. The derivatives were directly analyzed by MALDI-TOF-MS. Reaction conditions such as reaction temperature,concentration of acetic acid and excess ratio of CPH regent were optimized. When the oligosaccharides reacted with 100 time excess of CPH at 100 ℃ and in 0. 125% acetic acid （ V/V） ,the yield was highest. The results showed the derivatization with CPH increased the sensitivity of MS detection and this method was suitable for the analysis of oligosaccharides′ compositions.