固相萃取-串联液质法测定重组人粒细胞刺激因子注射液中氨苄青霉素残留

Solid-phase extraction LC-MS/MS determination of ampicillin residue in recombinant human granulocyte colony-stimulating factor injection

目的:建立SPE-LC-MS/MS法测定重组人粒细胞刺激因子注射液中氨苄青霉素残留。方法:选择阿莫西林为内标,样品经过HLB固相萃取小柱净化,浓缩,有效地去除杂质。采用Agilent Eclipse XDB-C18(4.6 mm×150 mm,5μm)色谱柱,以乙腈-水(甲酸调pH 3.1)为流动相进行梯度洗脱。采用电喷雾离子源(ESI),正离子模式,多反应监测(MRM)。氨苄青霉素,内标监测离子对的m/z分别为350.0/106.1,350.0/192.0和366.0/114.0,366.0/208.0。结果:氨苄青霉素浓度在0.0442～3.5362μg.L-1范围内线性关系良好(r=0.9995),最低定量限为0.0442μg.L-1,方法回收率在101.1%～106.7%之间,批内,批间RSD分别小于5.8%及7.3%。结论:本方法准确可靠,重复性好,灵敏度高,适用于重组人粒细胞刺激因子注射液中氨苄青霉素残留检测。

Objective：To establish a solid-phase extraction-LC-MS/MS method for the determination of ampicillin residue in recombinant human granulocyte colony stimulating factor（rhG-CSF）.Methods：Using amoxicillin as the internal standard,the sample was purified and concentrated by a HLB solid-phase extraction column.Chromatographic separation was performed on a Agilent Eclipse XDB-C18（4.6 mm×150 mm,5 μm）column with the mobile phase of acetonitrile-water（adjust pH to 3.1 with formic acid） by gradient elution.Electrospray ionization（ESI） source was applied and operated in positive mode,ampicillin and amoxicillin were detected on multiple reaction monitoring（MRM） mode by the transitions from the precursor to the production（m/z 350.0/106.1,350.0/192.0 and 366.0/114.0,366.0/208.0）.Results：For ampicillin,calibration curve was linear in range from 0.0442 to 3.5362 μg·L-1（r=0.9995）,the limit of quantitation was 0.0442 μg·L-1,the method recovery was between 101.1%～106.7%,the RSDs of inter-and intra-day precisions were less than 5.8% and 7.3% respectively.Conclusion：This method is be accurate,good reproducibility and sensitive for determination of ampicillin residue in rhG-CSF.