HPLC-DAD-MS法测定泮托拉唑钠原料及其制剂中的3种已知杂质

Determination of Pantoprazole Sodium and Its Three Impurities by HPLC-DAD-MS

目的建立高效液相色谱-二极管阵列检测器-质谱法（HPLC-DAD-MS）同时测定泮托拉唑钠及其3种微量杂质。方法采用RP-HPLC,色谱柱：Alltima C18柱（4.6 mm×250 mm,5μm）;流动相：乙腈-0.02 mol·L^-1醋酸铵梯度洗脱,梯度条件如下：0 min,30∶70;5 min,40∶60;20 min,40∶60;20.01 min,30∶70;30 min,30∶70;流速：0.8 mL.min-1;柱温：25℃;进样量：10μL;检测波长：288 nm;离子源：ESI,扫描模式：正离子扫描。结果泮托拉唑钠及其3种杂质分离良好。泮托拉唑钠、5-二氟甲氧基-2-{[（3,4-二甲氧基-2-吡啶基）-甲基]硫}-1H-苯并咪唑（中间体Ⅰ）、泮托拉唑砜、5-二氟甲氧基-2-[（3,4-二甲氧基-1-氧代-2-吡啶基）-甲磺酰基]-1H-苯并咪唑（副产物Ⅲ）分别在0.002-2.50,0.002-2.51,0.002-2.51,0.002-2.47μg内线性关系良好（r=0.999 9）;最低检测限分别为0.6,0.25,0.4,0.2 ng。结论该方法简便准确,可靠,重现性好,方法可行,可作为泮托拉唑钠及其3种可能微量杂质的质量控制方法。

OBJECTIVE To establisha a high performance liquid chromatography-diode array detection-mass spectrometry（HPLC-DAD-MS） assay for determining pantoprazole sodium and its three impurities simultaneously.METHODS The separation was achieved on a C18 column（4.6 mm×250 mm,5 μm）utilizing a mobile phase of acetonitrile-0.02 mol·L^-1 ammonium acetate（0 min,30∶70;5 min,40∶60;20 min,40∶60;20.01 min,30∶70;30 min,30∶70）at a flow rate of 0.8 mL·min^-1.The column temperature was set at 25 ℃.The compounds were monitored at 288 nm,and an ESI source was used with a positive ionization mode.RESULTS Pantoprazole sodium and its three impurities were determined by the provided method.Pantoprazole sodium,5-（difluoromethoxy）-2-{[（3,4-dimethoxy-2-py-ridinyl）methyl]thio}-1H-benzimidazole（intermediate I）,pantoprazole sulfone,5-（difluoro-methoxy）-2-[（3,4-dimethoxy-1-oxido-pyridinyl）-methyl sulfonyl]-1H-benzimidazole（by-product Ⅲ） were liner over the range of 0.002-2.50 μg（r=0.999 9）,0.002-2.51 μg（r=0.999 9）,0.002-2.51 μg（r=0.999 9）,0.002-2.47 μg（r=0.999 9）,respectively.The values of limit of detection were 0.6,0.25,0.4,0.2 ng for the four compounds.CONCLUSION Pantoprazole sodium and its three impurities can be separated with a good resolution.The method appeared to be simple,accurate,reliable and reproducible,and could be applied to control the quality of pantoprazole sodium.