色谱

色谱 ›› 2012, Vol. 30 ›› Issue (04): 395-399.

• 特别策划 • 上一篇    下一篇

高效离子交换色谱-电化学检测法测定药物合成中间体中的6-磷酸甘露糖

潘广文*, 刘绿叶, 叶明立, 胡忠阳   

  1. 赛默飞世尔科技有限公司上海实验室, 上海 201203
  • 收稿日期:2011-12-19 修回日期:2012-02-27 出版日期:2012-04-28 发布日期:2012-04-28
  • 通讯作者: 潘广文
  • 基金资助:

    安徽省教育厅高校省级科学研究项目(KJ2011B211)

Determination of mannose-6-phosphate by high performance ion-exchange Chromatography and electrochemical detector

PAN Guangwen*, LIU Lüye, YE Mingli, HU Zhongyang   

  1. Shanghai Lab., Thermo Fisher Scientific Inc., Shanghai 201203, China
  • Received:2011-12-19 Revised:2012-02-27 Online:2012-04-28 Published:2012-04-28
  • Contact: Pan Guang-Wen

PDF

324

被引次数

8

摘要： 在制备6-磷酸甘露糖过程中,将6-磷酸甘露糖与磷酸根杂质分开是纯化过程和建立质量标准的重要环节。本文建立了6-磷酸甘露糖和磷酸根的离子色谱分离-电化学检测方法。样品经溶解、过滤后进行色谱分析,以IonPac AG18离子交换柱(50 mm×4 mm)为保护柱,在Ionpac AS18离子交换色谱柱(250 mm×4 mm)上分离,以25 mmol/L氢氧化钾溶液为流动相,等度淋洗,流速1.0 mL/min,安培检测器和电导检测器串联检测,外标法定量。先使用安培检测器检测,在碱性条件下,6-磷酸甘露糖在安培检测器上被选择性检出;后使用电导检测器检测,经ASRS型抑制器抑制背景电导后,6-磷酸甘露糖和磷酸根同时被电导检测器检出,二者分离度良好。采用安培检测器检测时,进样量为25 μL, 6-磷酸甘露糖的线性范围为0.06～10.00 mg/L,相关系数为0.9998,回收率为92.1%～103.1%,相对标准偏差均小于3%,检出限(以信噪比为3计)为0.02 mg/L。该方法灵敏度高,无杂质干扰,前处理简便,可用于原料药合成中间体的检测,结果令人满意。

关键词: 6-磷酸甘露糖, 安培检测器, 串联检测, 电导检测器, 离子交换色谱, 药物合成中间体

Abstract: A new method was developed for the determination of mannose-6-phosphate （M6P）of intermediate by high performance ion-exclusion chromatography(IC). The sample was dissolved with purified water, filtrated by 0.22 μm nylon filter membrane, and then analyzed by Ion-exchange Chromatography. The analytical column was IonPac AG18 and IonPac AS18 (250 mm × 4 mm), potassium hydroxide was used as the eluent at a flow rate of 1.0 mL/min. The detection was performed with electrochemical detector (amperometric detector and conductive detector in line). The quantitive analysis of external standard calibration curves was used. With amperometric detector the linear range of the method for mannose-6-phosphate were 0.06 — 10.0 mg/L (r=0.9998). The average recoveries were 92% — 104% with RSDs less than 3%. The detection limit was 0.02 mg/L for mannose-6-phosphate. The method is simple, effective, sensitive and selective. It can be used to determine the quality of intermediate for chemosynthetic industry.

Key words: electrochemical detector, ion-exchange chromatography(IC), mannose-6-phosphate (M6P)