1-(4-异丙基)苯基-3-甲基-5-吡唑啉酮衍生化寡糖的HPLC-ESI/MS在线联用分析

Analysis of Oligosaccharides Labeled with 1-(4-Isopropyl) phenyl-3-methyl-5pyrazolone Using HPLC Coupled with On-line ESI-MS Techniques

为了给寡糖质量控制和分析研究提供方法参考,对1-(4-异丙基)苯基-3-甲基-5-吡唑啉酮(PPMP)衍生化试剂在液相色谱(HPLC)和电喷雾质谱(ESI-MS)技术分析寡糖中的应用进行研究。用PPMP试剂在氨水介质中对葡寡糖链进行衍生化,衍生化产物用RP-HPLC分离,ESI-MS分析。实验表明:在确定的衍生化条件下,PPMP和寡糖的衍生化主产物为单分子标记物,葡寡糖混合物的单分子PPMP衍生物在ESI-MS的正负离子模式下均有较好的响应,并且在RP-HPLC柱上能够实现很好地分离。据此建立了PPMP柱前衍生,HPLC-ESI/MS在线联用分析寡糖混合物组成的方法。该方法定性准确、样品处理简单、分析速度快、结果准确可靠,可作为寡糖样品的质量控制、构效关系研究等方面的一种测试方法。

Maltodextrin was derivatized with 1-（4-isopropyl） phenyl-3-methyl-5-pyrazolone （PPMP） in aqueous ammonia and derivatization products were analyzed by RP-HPLC sepa- ration and ESI-MS identification. The results indicated that mono-PPMP derivatives are still principal product for oligosaccharides. The mono-PPMP-derivatives for oligosaccharides from maltodextrins show satisfactory response to ESI-MS both in positive-ion mode and in negative-ion mode. The mixture of oligosaccharides derivatives with degree of poly- merization 1--17 was successfully separated by RP-HPLC. A new method for detecting the composition of oligosaccharides mixture with PPMP pre-column derivatization by HPLC coupled with on-line ESI-MS techniques was established. The method shows following merits. simple treatment for sample, fast analysis and accurate qualitation, and it can be used as a reference method for research of oligosaccharides＇ quality control and structure function relationship.