目的:建立一种超高效液相色谱-串联质谱联用法,用于测定注射用头孢美唑钠中的13个潜在亚硝胺类基因毒性杂质的含量(NDELA、NMOR、NPYR、NMPhA、NEPhA、NDiBA、NDBzA、NDBA、NDMA、NDEA、NPIP、NDiPA和NDPA)。方法:使用ACQUITY UPLC BEH C8(100 mm×2.1 mm,1.7 μm)色谱柱和Agilent Infinity Lab Poroshell120 Bonus-RP(100 mm×3.0 mm,2.7 μm)色谱柱,流动相为0.1%甲酸(A)-甲醇(B),梯度洗脱,流速0.4 mL·min-1,采用APCI离子源,在多反应监测模式(MRM)下,对全国20个生产厂家以及国外原研厂家的样品进行分析。结果:该方法能同时测定13个亚硝胺类基因毒性杂质。其线性范围、灵敏度、精密度、回收率均满足分析要求。在21批注射用头孢美唑钠中,10批检出了NPIP,但均小于定量限。结论:注射用头孢美唑钠中存在亚硝胺类基因毒性杂质的风险可控;所建立的方法高效灵敏,性能优异,专属性强,可为以四氮唑环为结构特征的抗生素中亚硝胺类基因毒性杂质控制方法提供参考,也可用于胶塞中N-亚硝胺类基因毒性杂质迁移量的检测,为监管部门提供技术储备和数据支持。
Objective: To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for the simultaneous determination of 13 potential nitrosamine genotoxic impurities (NDELA, NMOR, NPYR, NMPhA, NEPhA, NDiBA, NDBzA, NDBA, NDMA, NDEA, NPIP, NDiPA and NDPA) in cefmetazole sodium for injection. Methods: This assay was performed on chromatographic column ACQUITY UPLC BEH C8(100 mm×2.1 mm, 1.7 μm) and chromatographic column Agilent Infinity Lab Poroshell120 Bonus-RP(100 mm×3.0 mm, 2.7 μm) with mobile phase 0.1% formic acid (A)-methanol (B) by gradient elution at a flow rate of 0.4 mL·min-1. Multiple reaction monitoring (MRM) was performed on a triple quadripole mass spectrometer with an APCI source to analyze samples from 20 manufacturers in China and a foreign original manufacturer. Results: 13 kinds of nitrosamine genotoxic impurities were detected simultaneously by this method. The linear range, sensitivity, precision, and recovery of 13 compounds met the analysis requirements. Among 21 batches of cefmetazole sodium for injection, NPIP was detected in 10 batches, but all were less than the limit of quantification. Conclusion: The risk of nitrosamines genotoxic impurities in cefmetazole sodium for injection is controllable. The established method is efficient and sensitive, and has excellent performance and strong specificity. It can be used as a method for controlling nitrosamine genotoxic impurities in antibiotics with a tetrazolium ring as the structural feature. It can also be used for the detection of N-nitrosamine genotoxic impurities migration in rubber stoppers, and provide technical reserves and data support for regulatory authorities.
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