气相色谱-质谱联用法测定培美曲塞二钠起始原料中遗传毒性杂质甲磺酸乙酯
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摘要
目的建立测定培美曲塞二钠起始原料中的遗传毒性杂质甲磺酸乙酯含量的气相色谱-质谱联用(GC-MS)法。方法采用GCMS法,以N,N-二甲基甲酰胺为溶剂。用Agilent DB-624毛细管色谱柱(30 mm×0.32 mm,1.8μm),以高纯氦气为流动相,流速为3.0 m L/min(线速度为65 cm/s),柱温为120℃维持2 min,以15℃/min的速率升温至165℃;进样方式为分流进样,分离比为10∶1,进样口温度为150℃;离子源采用Electron Impact(EI)电子轰击源,扫描方式为选择离子扫描模式(SIM);检测器电压(相对值)为0 k V;接口温度为200℃,离子源温度为230℃;溶剂切除时间为2.5 min;进样量为2μL。采用标准曲线法进行定量。结果甲磺酸乙酯测定的专属性强,检测限为0.11μg/g(S/N=3),定量限为0.38μg/g(S/N=10);其质量浓度在7.99~159.84 ng/m L范围内与峰面积线性关系良好;平均加样回收率为97.94%,RSD为3.85%(n=9);精密度的RSD为8.52%(n=6),重复性的RSD为1.09%(n=6)。结论该方法专属性强,准确度高,适用于培美曲塞二钠起始原料中遗传毒性杂质甲磺酸乙酯的测定。
Objective To establish a GC-MS method for the determination of the genotoxic impurity ethyl methanesulfonate in starting material of Pemetrexed Disodium by GC-MS.Methods GC-MS was conducted,and the samples were dissolved by N,N-Dimethylformamide.The column was Agilent DB-624(30 mm × 0.32 mm,1.8 μm) capillary column,the inlet temperature was 150 ℃,column flow was 3.0 m L/min(linear velocity was 65 cm/s),carrier gas was high purity helium,column temperature was set at 120 ℃,hold for 2 min,then increased to 165 ℃ at 15 ℃/min,split injection,split ratio was 10 ∶ 1,the detector is a mass spectrometer detector with a electron impact ion source,ion source temperature was 230 ℃,scanning(detection) method was selective ion monitoring,detector voltage was 0 k V(respect to the tuning results),the interface temperature was 200 ℃,the delay time of solvent was 2.5 min,the injection volume was 2 μL,the concentration was calculated by standard curve.Results The specificity of the method was good,the LOD and LOQ were 0.11 μg/g(S/N = 3) and 0.38 μg/g(S/N = 10) in starting material.The calibration curve was linear in the range of7.99-159.84 ng/m L,the recoveries were 97.94%(n = 9),RSD was 3.85%(n = 6).RSD of precision was 8.52%(n = 6),and RSD of repeatability was 1.09%(n = 6).Conclusion The method has good specificity and high accuracy,which can be used to measure the genotoxic impurity ethyl methanesulfonate in starting material of Pemetrexed Disodium.
Objective To establish a GC-MS method for the determination of the genotoxic impurity ethyl methanesulfonate in starting material of Pemetrexed Disodium by GC-MS.Methods GC-MS was conducted,and the samples were dissolved by N,N-Dimethylformamide.The column was Agilent DB-624(30 mm × 0.32 mm,1.8 μm) capillary column,the inlet temperature was 150 ℃,column flow was 3.0 m L/min(linear velocity was 65 cm/s),carrier gas was high purity helium,column temperature was set at 120 ℃,hold for 2 min,then increased to 165 ℃ at 15 ℃/min,split injection,split ratio was 10 ∶ 1,the detector is a mass spectrometer detector with a electron impact ion source,ion source temperature was 230 ℃,scanning(detection) method was selective ion monitoring,detector voltage was 0 k V(respect to the tuning results),the interface temperature was 200 ℃,the delay time of solvent was 2.5 min,the injection volume was 2 μL,the concentration was calculated by standard curve.Results The specificity of the method was good,the LOD and LOQ were 0.11 μg/g(S/N = 3) and 0.38 μg/g(S/N = 10) in starting material.The calibration curve was linear in the range of7.99-159.84 ng/m L,the recoveries were 97.94%(n = 9),RSD was 3.85%(n = 6).RSD of precision was 8.52%(n = 6),and RSD of repeatability was 1.09%(n = 6).Conclusion The method has good specificity and high accuracy,which can be used to measure the genotoxic impurity ethyl methanesulfonate in starting material of Pemetrexed Disodium.
引文
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[12]冯慧敏,杭太俊,高新桃,等.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[J].药物分析杂志,2014,55(12):2202-2206.
[2]张慧敏,林建群,冯康彪,等.药品中遗传毒性杂质的评估和控制[J].中国现代应用药学,2014,31(9):1160-1166.
[3]马磊,马玉楠,陈震,等.遗传毒性杂质的警示结构[J].中国新药杂志,2014,23(18):2106-2111.
[4]王欣,王雪,宋捷,等.大鼠体内彗星试验方法的建立与应用研究[J].药物评价研究,2012,35(1):6-9.
[5]段佳,陈祥贵,帅培强,等.Hep G2和L5178Y细胞彗星试验检测甲磺酸甲酯和环磷酰胺遗传毒性的比较研究[J].西华大学学报:自然科学版,2011,30(1):19-20.
[6]王怡净.人类细胞体外多终点致突变试验系统的建立[D].成都:四川大学,2005.
[7]连小刚,李孝壁,程红艳.衍生气相色谱法测定甲磺酸伊马替尼中甲磺酸烷基酯类[J].中国医药科学,2013,3(17):128-130.
[8]王莉芳,郭文敏,杨汉煜,等.GC-MS法同时测定SIPI5357甲磺酸盐中3种甲磺酸酯[J].中国新药杂志,2014,23(11):1326-1329.
[9]刘立云,张倩如,孙连福,等.GC-MS法同时测定盐酸决奈达隆中的3种甲磺酸酯[J].中国药师,2014,17(8):1274-1276.
[10]邱妍川,李跃芬,钟玲,等.毛细管气相色谱法测定阿仑膦酸钠原料药中甲烷磺酸乙酯的残留量[J].中国药房,2014,25(24):2295-2297.
[11]杨茉,于润芳,王志凤.气相色谱法测定甲磺酸达比加群酯中的甲磺酸乙酯[J].继续医学教育,2014,28(9):42-44.
[12]冯慧敏,杭太俊,高新桃,等.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[J].药物分析杂志,2014,55(12):2202-2206.