UPLC-MS/MS法测定利奈唑胺原料和葡萄糖注射液中的叠氮类遗传毒性杂质

Determination of azide genotoxic impurity in linezolid API and glucose injection by UPLC-MS/MS

建立了一种超高效液相色谱-串联质谱法(UPLC-MS/MS)测定利奈唑胺原料及其葡萄糖注射液中的遗传毒性杂质(R)-5-(叠氮甲基)-3-[3-氟-4-(4-吗啉基)苯基]-2-唑烷酮。采用Acquity UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm)进行色谱分离,以0.1%甲酸水-0.1%甲酸乙腈(60∶40)为流动相,流速为0.3 mL/min进行等度洗脱;采用电喷雾离子化,在正离子模式下多反应监测模式进行监测。结果表明,该遗传毒性杂质在质量浓度4~12 ng/mL内线性关系良好,定量限为0.073 ng/mL,低、中、高浓度(80%、100%、120%限度浓度)加样溶液平均回收率分别为101.14%、100.59%和101.47%(RSD分别为0.73%、1.10%、0.91%)。同时,该遗传毒性杂质供试品溶液在6 d内稳定,3批制剂及2批原料中均未检出(R)-5-(叠氮甲基)-3-[3-氟-4-(4-吗啉基)苯基]-2-唑烷酮。

A UPLC-MS/MS method was established for the determination of the genotoxic impurity(R)-5-(azidomethyl)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxazolidinone in linezolid API and its glucose injection.Chromatographic separation was performed on a Waters Acquity UPLC HSS T3 column(100 mm×2.1 mm,1.8μm)with 0.1%formic acid water-0.1%formic acid acetonitrile(60∶40)at a flow rate of 0.3 mL/min.The UPLC-MS/MS was equipped with electrospray ionization in positive ionization mode and multiple reaction monitoring mode.The results showed that the calibration curve was linear in the range of 4-12 ng/mL and the limit of quantification was 0.073 ng/mL.The average recoveries of the low,medium and high concentration(80%,100%,120%limit concentration)loading solutions were 101.14%,100.59%and 101.47%,respectively(RSDs:0.73%,1.10%and 0.91%,respectively).The sample solution was stable for 6 d.No genotoxic impurity of(R)-5-(Azidomethyl)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxazolidinonewas not detected in the samples of linezolid API and its glucose injection.