中药水提液中还原糖衍生化测定方法研究  

Study on the method of reducing sugar derivatization in water-extract of Chinese materia medica

作　　者：田妍[1] 陈振兴[1] 张宁[1] 王翠玲[1] 刘竹兰[1] 刘建利[1] 

机构地区：[1]西部资源生物与现代生物技术省部共建教育部重点实验室,西北大学生命科学学院,西安710069

出　　处：《中华中医药杂志》2016年第9期3803-3807,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：长江学者和创新团队发展计划项目(No.IRT1174);国家自然科学基金项目(No.20872118,No.30070905);陕西省重点实验室基金项目(No.2010JS097,No.11JS090,No.12JS110);陕西省教育厅基金项目(No.12JK1010)~~

摘　　要：目的:建立1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱法测定中药水提液中还原糖种类及含量的方法。方法:以7种还原糖作为标准物,通过PMP柱前衍生化,利用反相高效液相色谱法,分析其衍生产物。以PMP衍生产物峰面积大小为指标,分别对衍生反应的反应时间、反应温度、PMP和Na OH用量、后处理使用的萃取试剂、萃取次数进行考察,得到了最佳衍生化条件。并将该方法用于7味中药水提液中还原糖类成分种类及含量的分析。结果:葡萄糖是本研究所检测7味中药中普遍存在的一类成分,且含量较高,除标准糖外,还含有未知还原糖类成分。D+(葡萄糖)、D+(半乳糖)、D+(甘露糖)、D+(麦芽糖)和D+(乳糖)、D+(阿拉伯糖)和D+(木糖)的线性范围分别为0.72-36μg/m L(r=0.9997)、0.72-36μg/m L(r=0.9998)、0.72-36μg/m L(r=0.9995)、1.44-72μg/m L(r=0.9995)、0.6-30μg/m L(r=0.9995)。结论:本方法简便、灵敏、重复性好,可用于中药水提液中还原糖类成份种类及含量的分析。Objective：To establish 1-pheny-3-methyl-5-Pyrazolone（PMP） precolumn derivation HPLC method to determine the types and contents of reducing suger in water-extract of Chinese materia medica.Methods：7 kinds of reducing sugars,as the standard substance,were performed by a PMP precolumn derivation,which would be analyzed with reversed-phase HPLC.With the peak area of PMP derivatives as the index,the reaction time and temperature in derivative reaction,the dosage of PMP and Na OH as well as the extraction reagents and times in post-processing using were respectively analyzed to get the optimal derivatization conditions.The method was used to analyze the types and contents of reducing carbohydrates in 7 kinds of waterextracts in Chinese materia medica.Results：Glucose was a ubiquitous composition in 7 Chinese materia medicas with a higher content than others.Besids the standard sugar,there were still some unknown reducing sugars in the Chinese materia medicas.The D＋（Glucose）,D＋（Galactose）,D＋（Mannose）,D＋（Maltose）,D＋（Lactose）,D＋（Arabinose） and D＋（Xylose） were a linearity range of 0.72-36μg/m L（r=0.9997）,0.72-36μg/m L（r=0.9998）,0.72-36μg/m L（r=0.9995）,1.44-72μg/m L（r=0.9995）,0.6-30μg/m L（r=0.9995）,respectively.Conclusion：With its simple,sensitive and repeatable merits,this method was suitable to analyze the types and contents of reducing sugar in water-extract of Chinese materia medica.

关 键 词：中药 还原糖 柱前衍生化 1-苯基-3-甲基-5-吡唑啉酮 高效液相色谱 

分 类 号：R284[医药卫生—中药学]