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  

Determination of mannose-6-phosphate in pharmaceutical intermediates by high performance ion-exchange chromatography-electrochemical detection

作　　者：潘广文 刘绿叶 叶明立 胡忠阳 

机构地区：[1]赛默飞世尔科技有限公司上海实验室,上海201203

出　　处：《色谱》2012年第4期395-399,共5页Chinese Journal of Chromatography

基　　金：安徽省教育厅高校省级科学研究项目(KJ2011B211)

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Ion-Pac AG18离子交换柱(50 m m×4 m m)为保护柱,在Io npac AS18离子交换色谱柱(250 m m×4 m m)上分离,以25 m m o l/L氢氧化钾溶液为流动相,等度淋洗,流速1.0 m L/m in,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25μL,6-磷酸甘露糖的线性范围为0.06～10.00 mg/L,相关系数为0.999 8,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 m g/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。A novel method was developed for the determination of mannose-6-phosphate and the phosphate in pharmaceutical intermediates by high performance ion-exchange chromatography（IC） and electrochemical detection.The sample was dissolved with purified water,filtrated by a 0.22 μm nylon filter membrane,and then separated by ion-exchange chromatography.The separation was performed on an IonPac AS18 column（250 mm×4 mm） with an IonPac AG18（50 mm×4 mm） as guard column,and 25 mmol/L KOH solution as the eluent at a flow rate of 1.0 mL/min.The detection was performed with electrochemical detectors（an integrated pulsed amperometric detector and a suppressed conductivity detector in line）.Mannose-6-phosphate was selectively determined with an integrated pulsed amperometric detector first.Then,both of mannose-6-phosphate and phosphate were determined with a suppressed conductivity detector after the background conductance of KOH was suppressed with the ASRS suppressor.The injection volume was 25 μL.The external standard calibration curve was used for quantitative analysis.With an amperometric detector,the linear range of the method for mannose-6-phosphate were 0.06-10.0 mg/L（r=0.999 8）.The recoveries were 92.1%-103.1% with the relative standard deviations（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 pharmaceutical intermediates.

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

分 类 号：O658[理学—分析化学]