建立了亲水/反相二维色谱用于制备桔梗中三萜皂苷单体的方法。桔梗经水煮醇沉、反相和亲水两种模式的固相萃取后得到三萜皂苷类组分。选定 XAmide色谱柱（150 mm×20 mm，5μm），以乙腈和水为流动相，在亲水色谱模式下进行组分制备。选择时间触发模式，以1 min为单位进行馏分收集，得到6~25 min 之间的20个三萜皂苷精细组分。以第18个馏分（JG23）为例，在反相色谱模式下采用 Atlantis Prep T3色谱柱（100 mm×30 mm，5μm）制备，得到两个单体化合物。通过质谱和核磁共振对其进行定性，确定分别为deapi-platycoside E和platyco-side E。实验结果表明，该制备方法具有好的正交选择性，对于复杂样品中三萜皂苷类化合物的分离纯化有一定的借鉴意义。
A two-dimensional(2-D)preparative liquid chromatography method was developed for the preparation of triterpene saponins from Platycodon grandiflorum using hydrophilic interaction liquid chromatography( HILIC)coupled with reversed phase liquid chromatography ( RPLC). At first,the crude extract was obtained from Platycodon grandiflorum by boiled alco-hol precipitation. Then,the concentrated crude extracts were continuously pretreated using sol-id phase extraction( SPE)under reversed-phase and hydrophilic-phase modes to remove the impurities. Subsequently,XAmide column(150 mm×20 mm,5 μm)was selected to separate the triterpene saponin constituents under HILIC mode using water and acetonitrile as mobile phases. From 6 min to 25 min,each fraction was collected per minute under time-triggered mode and 20 fractions were collected. The 18th fraction( JG23)was selected for further purifi-cation. The column of Atlantis PrepT3(100 mm×30 mm,5 μm)was chosen and two monomer-ic compounds were obtained. The two compounds with over 90% purity were identified as dea-pi-platycoside E and platycoside E with mass spectrometry( MS)and nuclear magnetic reso-nance( NMR). This 2-D HILIC-RPLC method with high orthogonality can be used in the prepa-ration of triterpene saponins from natural products.
参考文献
| [1] | Zhao X L. China Condiment(赵秀玲. 中国调味品),2012, 37(2):5 |
| [2] | He Z D,Qiao C F,Han Q B,et al. Tetrahedron,2005,61:2211 |
| [3] | He M L,Cheng X W,Chen J K,et al. Traditional Chinese Medicine New Drug and Clinical Pharmacology(何美莲,程小卫,陈家宽,等. 中药新药与临床药理),2005,16(6):457 |
| [4] | Young W H,Yeong S K. Phytochem Anal,2009,20:207 |
| [5] | In J H,Minseok K,Yun C N,et al. J Sep Sci,2011,34:2559 |
| [6] | Zhu F X,Jia X B,Cai Y,et al. Acta Chromatographica, 2013,25(1):147 |
| [7] | Sun B S,Gu L J,Fang Z M,et al. J Pharm Biomed Anal, 2009,50:15 |
| [8] | Zhang J,Wang L L,Shan L G,et al. Chinese Journal of Chromatography(张静,王玲玲,单联国,等. 色谱),2012, 30(8):804 |
| [9] | Shi Y M,Yang H,Li Y Y,et al. Food Research and Develop-ment(史一鸣,杨虹,李燕艳,等. 食品研究与开发),2012,33(12):134 |
| [10] | Noel S Q,Nerissa L D,Azamjon B S,et al. Anal Bioanal Chem,2007,389:1477 |
| [11] | Xing Q Q,Liang T,Shen G B,et al. Analyst,2012,137:2239 |
| [12] | Boguslaw B,Sylwia N. Anal Bioanal Chem,2012,402:231 |
| [13] | Li W,Zhang W,Xiang L,et al. Molecules,2010,15:8702 |
- 下载量()
- 访问量()
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%