色谱

色谱

• 研究论文 • 上一篇    下一篇

1-(2-萘基)-3-甲基-5-吡唑啉酮衍生试剂的制备及其在高效液相色谱-质谱法测定糖类化合物中的应用

孙志伟1,刘凌君1,户宝军1,盛筱1,王小艳2,索有瑞2,尤进茂1,2

  

  1. 1.Key Laboratory of Life-Organic Analysis of Sandong Province, College of Chemistry Science, Qufu Normal University, Qufu 273165, China; 2.Northwest Plateau Institute of Biology, the Chinese Academy of Sciences, Xining 810001, China
  • 收稿日期:2007-09-11 修回日期:2007-12-10 出版日期:2008-03-30 发布日期:1984-06-25
  • 通讯作者: 尤进茂

Preparation of 1-(2-naphthyl)-3-methyl-5-pyrazolone as pre-column derivatization reagent for the determination of saccharides using high performance liquid chromatography-mass spectrometry

SUN Zhiwei1, LIU Lingjun1, HU Baojun1, SHENG Xiao1, WANG Xiaoyan2, SUO Yourui2, YOU Jinmao1,2   

  1. 1.Key Laboratory of Life-Organic Analysis of Sandong Province, College of Chemistry Science, Qufu Normal University, Qufu 273165, China; 2.Northwest Plateau Institute of Biology, the Chinese Academy of Sciences, Xining 810001, China
  • Received:2007-09-11 Revised:2007-12-10 Online:2008-03-30 Published:1984-06-25

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摘要：

以1-(2-萘基)-3-甲基-5-吡唑啉酮(NMP)作为柱前衍生试剂,建立了简单、灵敏的糖类组分的反相高效液相色谱测定方法。NMP与糖在氨为催化剂的条件下,于70 ℃下反应可获得稳定的衍生产物。在Hypersil ODS 2反相色谱柱上,实现了8种单糖的基线分离。衍生物线性相关系数均大于0.9985,检出限为0.58～1.1 pmol。利用柱后在线串联质谱的电喷雾电离正离子模式监测，获得了各组分的质谱定性及裂解规律,特别是m/z 473的特征碎片离子可作为单糖NMP衍生物的判定依据。与1-苯基-3-甲基-5-吡唑啉酮(PMP)相比,NMP对糖的衍生化具有灵敏、简单、质谱裂解规律性强、重现性好等优点。该方法用于测定油菜花粉多糖中的单糖组成,结果令人满意。

关键词: 1-(2-萘基)-3-甲基-5-吡唑啉酮, 单糖 , 高效液相色谱-质谱, 柱前衍生

Abstract:

Eight saccharides were derivatized using 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) as pre-column derivatizing reagent， and separated on a reversed-phase Hypersil ODS 2 column (4.6 mm×200 mm, 5 μm), by high performance liquid chromatography (HPLC) in conjunction with a gradient elution, detected by a diode array detector (DAD), and identified by electrospray ionization-mass spectrometry (ESI-MS) in positive ion mode. NMP reacted with reductive saccharides easily in the presence of 17% ammonia water at 70 ℃. All linear correlation coefficients for saccharide derivatives were over 0.9985. The detection limits （at signal-to-noise of 3∶1） were 0.58-1.1 pmol for saccharide derivatives. The characteristic fragment ions, especially m/z 473, from the cleavage of NMP-labeled saccharides exhibited high regularity for the identification of the composition of saccharide mixture. The established method is sensitive and repeatable for the determination of saccharides.

Key words: 1-(2-naphthyl)-3-methyl-5-pyrazolone, saccharides , high performance liquid chromatography-mass spectrometry (HPLC-MS): pre-column derivatization