LC-MS法同时检测甲磺酸伊马替尼中3个磺酸酯基因毒杂质

Determination of potential carcinogenic and genotoxic sulfonate impurities in imatinib mesylate by a highly sensitive liquid chromatography-mass spectrometry method

目的：建立LC-MS法对甲磺酸伊马替尼原料中的3个磺酸酯基因毒杂质—甲磺酸甲酯（MMS）、甲磺酸乙酯（EMS）、甲磺酸异丙酯（IPMS）进行定量检测。方法：选用Agilent Poroshell 120 SB-C18色谱柱（2.7μm,4.6 mm×100 mm）,以10mmol·L^-1甲酸铵（含0.1%甲酸）为流动相A,甲醇为流动相B,梯度洗脱[0 min（80%A-20%B）→5 min（10%A-90%B）→10 min（10%A-90%B）→10.5 min（80%A-20%B）→18.5 min（80%A-20%B）],流速为0.4 m L·min^-1。ESI正离子化SIM模式下对3种磺酸酯进行同时检测,雾化氮气压力0.3 MPa、温度300℃、流速10 L·min^-1,毛细管电压3 k V,传输电压30 V。结果：MMS在10-100 ng·m L^-1内线性关系良好（r=0.9988）,定量限为10 ng·m L^-1;EMS在5-50 ng·m L^-1内线性关系良好（r=0.9992）,定量限为5 ng·m L^-1;IPMS在10-100 ng·m L^-1内线性关系良好（r=0.9993）,定量限为10 ng·m L^-1。MMS、EMS、IPMS的回收率分别为99.86%、101.7%、103.8%。结论：经方法学验证,该法可用于甲磺酸伊马替尼中3个磺酸酯基因毒杂质的同时检测,能为质量控制提供参考。

Objective：To determine three sulfonic acid esters, methyl methanesulfonate （MMS）, ethyl methane- sulfonate （EMS）, and isopropyl methanesulfonate （IPMS）, known as potential carcinogenic and genotoxic impuri- ties in imatinib mesylate by liquid chromatography -mass spectrometry （LC -MS）. Methods： The separation was carried out on a Poroshell SB - C18 column （2.7 μm, 4.6 mm × 100 mm） with the mobile phase consisting of 10 mmol · L^-1 ammonium formate containing 0. 1% （V/V） formic acid （A） and methanol （B） by gradient elution [0 min（ 80% A - 20% B ） →5min （ 10% A - 90% B ） →10 min （ 10% A - 90% B ） →10.5 min （ 80% A - 20% B ） → 18.5 rain （80% A -20% B） ] at a flow rate of 0.4mL · min^-1. The results of quadrupole mass spectrometer was equipped with an ESI （in the positive mode）. The conditions were as follows： nitrogen drying gas with nebulizer pressure of 0.3 MPa, the flow rate of 10 L · min^- 1 , the temperature of 300 ℃, the capillary voltage of 3 kV and the fragmentation voltage of 30 V. Results： The results of limit of quantification （LOQ） of MMS, EMS and IPMS were 10 ng · mL^-1, 5 ng · mL^-1 and 10 ng · mL^-1, respectively. Recoveries of MMS, EMS and IPMS were 99.86%, 101.7% and 103.8%, respectively. The linear calibration curves of MMS and IPMS were obtained over the concentration range of 10 - 100 ng · mL^-1, while the curve of EMS over 5 -50 ng · mL^-1. Conclusion： The methodology validation showed that this LC -MS method is useful as a quality control method for the simultaneous trace analysis of MMS, EMS and IPMS in imatinib mesylate.