摘要
目的:建立LC-MS/MS法检测替格瑞洛中基因毒性杂质4,6-二氯-2-(丙硫基)-5-氨基嘧啶(DCPA)和5-硝基-2-(丙硫基)嘧啶-4,6-二醇(DHPN)。方法:采用Ultimate?UHPLC XB-Phenyl色谱柱(100 mm×2.1 mm,1.8μm),以0.1%甲酸溶液为流动相A,0.1%甲酸的甲醇溶液为流动相B,梯度洗脱,流速0.3 mL·min^(-1),柱温为40℃;采用AB SCIEX ExionLC-QTRAP5500三重四极杆串联离子阱液质联用仪进行检测,电喷雾离子源(ESI~+)、多反应监测模式(MRM),定量离子对为m/z 240.1→197.8(DCPA)、m/z 232.2→213.9(DHPN)。结果:DCPA质量浓度在0.10~16.1 ng·mL^(-1)范围内与峰面积呈良好线性关系(r=0.9999);检测限为0.03 ng·mL^(-1),定量限为0.10 ng·mL^(-1),平均加样回收率为97.5%(RSD=9.9%,n=9);DHPN质量浓度在0.20~16.1 ng·mL^(-1)范围内与峰面积呈良好线性关系(r=0.9995),检测限为0.06 ng·mL^(-1),定量限为0.20 ng·mL^(-1),平均加样回收率为98.1%(RSD=1.5%,n=9)。重复性和中间精密度实验中,DHPN在100%限度浓度加标供试品溶液(n=12)中的加样平均含量为7.97μg·g^(-1),RSD=4.9%;供试品溶液(n=12)中DCPA的平均含量为0.94μg·g^(-1),RSD=4.3%,表明精密度良好。对照品溶液、供试品溶液、加标供试品溶液中DCPA和DHPN室温放置24 h内稳定。3批替格瑞洛样品(批号Q19010701G、Q19010702G、Q19010703G)中均未检测到DHPN,DCPA的含量依次为1.2、1.1和1.1μg·g^(-1),均符合规定。结论:该方法简便、准确、可靠,可用于替格瑞洛中DCPA和DHPN的检测。
Objective:To establish an LC-MS/MS analytical method for the detection of 4,6-dichloro-2-(propylthio)-5-aminopyrimidine(DCPA)and 5-nitrol-2-(propylthio)pyrimidine-4,6-diol(DHPN)of genotoxic impurities in ticagrelor.Methods:The separation was performed on a Ultimate?UHPLC XB-Phenyl column(100 mm×2.1 mm,1.8μm)with mobile phase consisting of 0.1%formic acid solution(mobile phase A)and 0.1%formic acid methanol solution(mobile phase B)by gradient elution at a flow rate of0.3 m L·min^(-1).The column temperature was 40℃.AB SCIEX ExionLC-QTRAP5500 triple quadrupole tandem ion trap liquid mass spectrometer was used for detection,electrospray ion source(ESI+),multiple reaction monitoring mode(MRM).The quantified ion pairs were m/z 240.1→197.8(DCPA)and m/z 232.2→213.9(DHPN).Results:The calibration curve was in a good linearity between the range of the concentration was0.10-16.1 ng·m L^(-1)for DCPA and peak area(r=0.9999).The limit of detection was 0.03 ng·m L^(-1)and the limit of quantification was 0.10 ng·m L^(-1);the average recovery was 97.5%(n=9),with RSD of 9.9%.The calibration curve was in a good linearity between the range of the concentration was 0.20-16.1 ng·m L^(-1)for DHPN and peak area(r=0.9995).The limit of detection was 0.06 ng·m L^(-1)and the limit of quantification was 0.20 ng·m L^(-1).The average recovery was 98.1%(n=9),with RSD of 1.5%.In repeatability and intermediate precision experiments,the average concentration of DHPN was 7.97μg·g^(-1)in the 100%limit concentration spiked solutions(n=12),with RSD of 4.9%;the average concentration of DCPA was 0.94μg·g^(-1)in the test solutions,with RSD of 4.3%,indicating good precision.At room temperature,DCPA and DHPN were stable within24 h,in the control solution,test solution and spiked test solution.In three batches of ticagrelor samples(batch number:Q19010701G,Q19010702G and Q19010703G),DHPN wasn’t detected,and the contents of DCPA were1.2μg·g^(-1),1.1μg·g^(-1)and 1.1μg·g^(-1),respectively,which all met the requirements.Conclusion:The m
作者
王庆鹏
骆献丽
陈晓英
李原
张明辉
刘山崎
WANG Qing-peng;LUO Xian-li;CHEN Xiao-ying;LI Yuan;ZHANG Ming-hui;LIU Shan-qi(Lepu Pharmaceuticals,Inc.,zhengzhou 450001,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2022年第10期1822-1830,共9页
Chinese Journal of Pharmaceutical Analysis
