目的: 建立了气相色谱-串联质谱(GC-MS/MS)检测盐酸二甲双胍制剂中N-亚硝基二甲胺(NDMA)含量的方法。方法: 采用100%聚乙二醇(PEG-20M)为固定液的毛细管柱(Agilent VF-WAXms,30 m×0.25 mm,0.25 μm),程序升温进行分离。质谱采用电子轰击电离源(EI)电离,正离子模式,选择离子监测(MRM)模式检测。NDMA定量离子对为m/z 74.0→44.0,碰撞能量(CE)为8 eV,定性离子对为m/z 74.0→42.0,碰撞能量(CE)为18 eV。结果: 该法专属性良好,溶剂和辅料对NDMA测定无干扰。NDMA在0.296~59.2 ng·mL-1质量浓度范围内线性关系良好,检测限(LOD)和定量限(LOQ)分别为0.189 ng·mL-1和0.592 ng·mL-1,精密度、稳定性试验满足检测要求,加标回收率为91.9%~101.0%,RSD均小于2.5%。使用该方法对67批盐酸二甲双胍制剂中的NDMA进行测定,结果30批检出了NDMA,其中15批盐酸二甲双胍制剂所含NDMA超过了限度,且全部为缓释片。同时,对部分缓释片原料药的NDMA含量进行了测定,结果从原料药到制剂NDMA含量有明显增量。结论: 本文建立的方法准确、快速、灵敏、专属性强,可用于盐酸二甲双胍制剂中遗传毒性杂质NDMA的含量测定。建议企业不仅对盐酸二甲双胍制剂原料药进行质量控制,也应对生产工艺进行评估,降低因工艺造成NDMA的增加。
Objective: To establish a method for the determination of N-nitrosodimethylamine (NDMA) in metformin hydrochloride by gas chromatography tandem mass spectrometry (GC-MS/MS). Methods: A capillary column with 100% polyethylene glycol (PEG-20M) (Agilent VF-WAXms,30 m×0.25 mm,0.25 μm)as stationary liquid was used for separation. The mass spectrum was detected by EI ionization, positive ion mode and MRM mode. The quantitative ion pair of NDMA was m/z 74.0→44.0, and the collision energy (CE) was 8 eV. The qualitative ion pair was m/z 74.0→42.0, and the collision energy (CE) was 18 eV. Results: The specificity of the method was good, and the solvents and excipients did not interfere with the determination of NDMA. The linear range of NDMA was 0.296-59.2 ng·mL-1, the limit of detection (LOD) was 0.189 ng·mL-1 and the limit of quantitation (LOQ) was 0.592 ng·mL-1. The precision and stability tests met the detection requirements. The recoveries were 91.9%-101.0%, and RSDs were less than 2.5%. The method was used to determine NDMA in 67 batches of metformin hydrochloride preparations. NDMA were detected from 30 batches. Among them, 15 batches exceeded standard, and all were sustained-release tablets. The analysis of the corresponding active pharmaceutical ingredients (APIs) of some sustained-release tablets showed that the NDMA content of the sustained-release tablets increased significantly from the APIs to the preparations. Conclusion: This method is accurate, rapid, sensitive and specific, which can be used to control the content of NDMA in metformin hydrochloride. Enterprises should not only pay attention to the quality control of APIs, but also conduct process evaluation to reduce the increase of NDMA.
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