高效液相色谱-电喷雾质谱联用分析PPMP衍生化的壳寡糖

以1-(4-异丙基)苯基-3-甲基-5-吡唑啉酮(PPMP)为衍生化试剂在氨水介质中对壳寡糖链进行衍生化,衍生化产物用RP-HPLC分离和ESI-MS分析。结果表明在确定的衍生化条件下,PPMP和壳寡糖的衍生化产物主要为单分子衍生物,此单分子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柱上能够实现很好的分离。据此建立了PPMP柱前衍生HPLC/ESI-MS在线联用检测壳寡糖混合物组成的方法。该法可作为壳寡糖样品在质量控制、构效关系研究等方面的方法参考。

Analysis of chitooligosaccharides labeled with 1-(4-isopropyl) phenyl-3-methyl-5-pyrazolone using HPLC coupled with on-line ESI-MS technique

Chitooligosaccharides were derivatized with 1-(4-isopropyl) phenyl-3-methyl-5-pyrazolone(PPMP) in aqueous ammonia and derivatization products were analyzed by RP-HPLC separation and ESI-MS identification.The results indicated that mono-PPMP-derivatives are still principal products for chitooligosaccharides and the mono-PPMP-derivatives show satisfactory response to ESI-MS both in positive-ion mode and in negative-ion mode.The mixture of mono-PPMP-labeled chinooligosaccharides containing degree of polymerization 1～17 was successfully separated by RP-HPLC.On these grounds,a new method used for detection of the composition of chinooligosaccharides mixture was established with PPMP pre-column derivatization by HPLC coupled with on-line ESI-MS technique.This method has both high performance separation of HPLC and high sensitivity of ESI-MS.It shows some merits as following:sample treatment is simple,analysis is fast and qualitation is precise,and the method can be taken as a reference method for research of chinooligosaccharides’ quality control and structure-function relationship.