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。
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 specificity,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.
作者
孙春艳
纪颖鹤
秦昆明
高珣
赵龙山
SUN Chunyan;JI Yinghe;QIN Kunming;GAO Xun;ZHAO Longshan(School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China;Yangtze River Pharmaceutical Group Co.,Ltd,Taizhou 225300,China;School of Pharmacy,Jiangsu Ocean University,Lianyungang 222001,China)
出处
《色谱》
CAS
CSCD
北大核心
2019年第12期1297-1304,共8页
Chinese Journal of Chromatography
关键词
高效液相色谱-串联质谱
基因毒性杂质
吉非替尼
high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)
genotoxic impurity
gefitinib
