天然产物研究与开发 ›› 2015, Vol. 27 ›› Issue (7): 1205-1209.doi: 10.16333/j.1001-6880.2015.07.015

• 研究简报 • 上一篇    下一篇

二维制备高效液相色谱分离塞隆骨强极性多肽

江磊1,4,赵换2,陶燕铎1,王硕1,4,邵赟1,王启兰1,张耀洲3*,梅丽娟1*   

  1. 1 中国科学院西北高原生物研究所,西宁 810001;2 瀚盟生物技术(天津)有限公司;3 天津大学药学院,天津 300072;4中国科学院大学,北京 100049
  • 出版日期:2015-07-20 发布日期:2015-07-23

Separation of High-Polarity Peptides Using Two dimensional Preparative HPLC

JIANG Lei1,4,ZHAO Huan2,TAO Yan-duo1,WANG Shuo1,4,SHAO Yun1,WANG Qi-lan1,ZHANG Yao-zhou3*,MEI Li-juan1*   

  1. 1 Key laboratory of Tibetan medicine research,Northwest Plateau Institute of Biology,Chinese Academy of Sciences,Xining 810001,China; 2 Harmonia Biotechnology (Tianjin) Limited Company,Tianjin University;  3 School of Pharmaceutical Science and Technology,Tianjin University,Tianjin 300072,China;  4 University of Chinese Academy of Sciences,Beijing 100039,China
  • Online:2015-07-20 Published:2015-07-23

摘要: 本实验建立了一种新型的利用高正交性的二维制备型高效液相色谱系统分离强极性动物药多肽的方法。本文以塞隆骨水提取物为研究对象,以亲水性C18 AQ制备型高效色谱柱为第一维分离柱,首先在一维分离中将目标混合物分成若干组份;然后以C18 MP制备型高效液相色谱柱为第二维色谱分离柱,将第一维分离后得到的组份纯化为单体化合物。本研究最终得到5个塞隆骨单体化合物,化合物纯度均超过98%。经Nano-LC-ESI-MS/MS鉴定和搜库分析,这些多肽的序列分别为:KTAILVKE、RGAPQDQE、LVGPGAPGR、GFAGD和KPQWHP。此研究方法速度快、效率高且重复性好,可以对类似的研究提供借鉴。

关键词: 高效液相色谱, 塞隆骨, 多肽, 二维分离

Abstract: In the present separation work,Osteon Myospalacem Baileyi water extracts were taken as an example to explain the purification method.The first dimensional preparation was carried out on a C18 AQ prep column to obtain fractions from the crude sample.A C18 MP prep column was used for the second dimensional preparation to purify single compounds from the fractions.The purities of all the compounds isolated from the fractions were higher than 98%,which indicated that the developed method was efficient to isolate single compounds with high purity from the extract of Osteon Myospalacem Baileyi.After structural identification by UPLC-LTQ-MS/MS,the sequences of the purified peptides were determined to be KTAILVKE,RGAPQDQE,LVGPGAPGR,GFAGD and KPQWHP,respectively.The developed method showed great potential in the purification of traditional animal medicines.

Key words: high performance liquid chromatography, Osteon Myospalacem Baileyi, peptides, two-dimensional separation

中图分类号: 

R966