色谱

色谱 ›› 2019, Vol. 37 ›› Issue (12): 1297-1304.DOI: 10.3724/SP.J.1123.2019.07002

• 研究论文 • 上一篇    下一篇

液相色谱-串联质谱法测定吉非替尼中4种基因毒性杂质

孙春艳1,2, 纪颖鹤1, 秦昆明3, 高珣3, 赵龙山1   

  1. 1.沈阳药科大学药学院, 辽宁 沈阳 110016;
    2.扬子江药业, 江苏 泰州 225300;
    3.江苏海洋大学药学院, 江苏 连云港 222001
  • 收稿日期:2019-07-02 出版日期:2019-12-08 发布日期:2020-12-11
  • 通讯作者: 高珣.Tel:+86-523-43520039,E-mail:gaoxun-509@163.com;赵龙山.Tel/Fax:+86-24-43520571,E-mail:longshanzhao@163.com.

A high-performance liquid chromatography-tandem mass spectrometry method for the quantitative determination of four genotoxic impurities in gefitinib

SUN Chunyan1,2, JI Yinghe1, QIN Kunming3, GAO Xun3, ZHAO Longshan1   

  1. 1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China;
    2. Yangtze River Pharmaceutical Group Co., Ltd, Taizhou 225300, China;
    3. School of Pharmacy, Jiangsu Ocean University, Lianyungang 222001, China
  • Received:2019-07-02 Online:2019-12-08 Published:2020-12-11

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摘要： 建立了一种同时测定吉非替尼中4种基因毒性杂质3-氯-4-氟苯胺、3，4-二氟苯胺、3-氟-4-氯苯胺和3，4-二氯苯胺的高效液相色谱-串联质谱（HPLC-MS/MS）方法。用Inertsil ODS-3柱（100 mm×3.0 mm，3μm）为色谱柱，以0.1%（体积分数，下同）甲酸水溶液-0.1%甲酸乙腈溶液为流动相，在电喷雾正离子模式下进行测定。该方法在特异性、线性、精密度、准确性、稳定性和耐用性方面得到了验证。4种基因毒性杂质在0.6~96.0μg/L范围内与峰面积呈良好线性关系。检测限和定量限分别为0.2~2.0μg/L和0.6~6.0μg/L。所有杂质的回收率为91.0%~98.5%。检测后，在批号16052301和R16052501-1样品中仅检测到3-氯-4-氟苯胺，但低于杂质限度（6 mg/L）。该方法简便可靠，可用于吉非替尼中4种基因毒性杂质的测定，并为质量控制提供参考。

关键词: 高效液相色谱-串联质谱, 基因毒性杂质, 吉非替尼

Abstract: In this work, a method for the determination of the amounts of the four genotoxic impurities in gefitinib has been developed. A simple and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) approach has been developed, optimized, and validated. The genotoxic impurities were 3-chloro-4-fluoroaniline, 3,4-difluoroaniline, 3-fluoro-4-chloroaniline, and 3,4-dichloroaniline. Separation was achieved on an Inertsil ODS-3 column (100 mm×3.0 mm, 3 μm). The column temperature was 40 ℃, and the running time was 12 min. A triple quadrupole mass detector with positive electrospray ionization in the multiple reaction monitoring (MRM) mode was applied. The method was validated in terms of its specifi-city, linearity, precision, accuracy, stability, and robustness. The correlation coefficient of each impurity was higher than 0.999 in the range of 0.6-96.0 μg/L. The limits of detection (LODs) and limits of quantity (LOQs) were in the ranges of 0.2-2.0 μg/L and 0.6-6.0 μg/L, respectively. The recoveries of all impurities at 6, 30, and 60 μg/L were within 91.0%-98.5%. All impurities were stable within 2 h. After detection, only 3-chloro-4-fluoroaniline was detected in the batch number 16052301 and R16052501-1 gefitinib samples, but its concentration was below the impurity limit (6 mg/L). This method is simple, reliable, and suitable for the determination of four genotoxic impurities in gefitinib. It can be further applied as a reference for quality control.

Key words: high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), genotoxic impurity, gefitinib

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