1-苯基-3-甲基-5-吡唑啉酮糖衍生物在反相高效液相色谱法分离中保留行为的探讨  

Discussion of the retention behavior of PMP saccharide derivatives in their RP-HPLC separation

作　　者：郭怀忠[1,2] 董露[1] 杨飞飞[1] 朱星娜 

机构地区：[1]河北大学药学院,河北保定071002 [2]药物化学与分子诊断教育部重点实验室,河北保定071002

出　　处：《中南药学》2017年第4期511-514,共4页Central South Pharmacy

基　　金：河北省自然科学基金(No.H2016201221);国家自然科学基金(No.21575033)

摘　　要：本文针对反相HPLC分离中,1-苯基-3-甲基-5-吡唑啉酮(PMP)先于PMP糖衍生物(PMP-SD)出峰的现象,通过控制降解β-环糊精得到系列寡糖,PMP衍生后采用HPLC法对其保留行为进行考察,并结合相关文献以及不同条件下PMP和PMP-SD的结构进行讨论,来说明其极性变化的原因。结果表明,吡唑啉酮环为烯醇结构时是PMP的优势构象,烯醇基解离使PMP极性显著增大。衍生糖上顺式结构的C-2、C-3位羟基仅可与一个PMP吡唑啉酮环上的酮基通过氢键环合,抑制酮基向烯醇基的转变,使衍生糖上的PMP极性降低。衍生糖上反式结构的C-2、C-3位羟基可与2个PMP上的酮基通过氢键环合,使衍生糖上PMP的极性进一步降低。在使用高盐浓度的流动相对PMP-SD的洗脱中,糖链的高极性基本不发挥作用。本文为糖类成分反相HPLC分离时色谱柱的选择和更有针对性的条件优化提供参考。For the phenomenon of derivatization reagent 1-phenyl-3-methyl-5-pyrazolone （PMP） was eluted prior to PMP saccharide derivatives （PMP-SD） in their reversed phase HPLC separation, we discussed the retention behavior through a series of oligosaccharides of partly degraded fl-cyclodextrin which was derived with PMP, then were analyzed by HPLC. The reason for the polarity change of PMP and PMP-SD was explained through the discussion on their structures under different conditions and related literatures. The results showed that the enol structure was the dominant conformation of pyrazolone ring in PMP. The polarity of PMP increased significantly when enol group was dissociated. Saceharide derivatives with cis-C-2, C-3-hydroxyl groups contained only a ring with ketone group of pyrazolone ring through hydrogen bond, The formation of the ring restrained the ketone group from transforming into enol group, and the polarity of PMP in these saccharide derivatives was reduced. Saecharide derivatives with trans-C-2, C-3-hydroxy groups contained two rings with ketone groups through hydrogen bond, and the polarity of PMP in these saccharide derivatives was further reduced. The polarity effect of saccharide chains didn＇t work when eluted by the mobile phase containing high concentration of salt. This study can provide reference for the selection of column and the targeted optimization of the chromatographic conditions when the saccharides are separated by reversed phase HPLC.

关 键 词：糖 1-苯基-3-甲基-5-吡唑啉酮 衍生物 反相高效液相色谱 极性 

分 类 号：O657.72[理学—分析化学]