5种β受体拮抗剂类药物中的N-亚硝基类杂质的含量研究  

Content study of N-nitroso impurities in 5 types ofβ-blockers

作　　者：田珩[1] 杨仪雪 戴聪 刘亚雄[1] 严全鸿[1] TIAN Heng;YANG Yixue;DAI Cong;LIU Yaxiong;YAN Quanhong(Guangdong Institute for Drug Control/NMPA Key Laboratory for Quality Control and Evaluation of Pharmaceutical Excipients,Guangzhou 510180,China)

机构地区：[1]广东省药品检验所/国家药品监督管理局药用辅料质量控制与评价重点实验室,广州510180

出　　处：《中国药房》2024年第8期936-941,共6页China Pharmacy

基　　金：广东省医学科学技术研究基金项目(No.B2023406)。

摘　　要：目的测定普萘洛尔、美托洛尔、阿替洛尔、艾司洛尔、比索洛尔原料药/制剂中N-亚硝基类杂质含量,明确其含量的关注阈值。方法采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱技术。以ACE Excel 3 C18-AR为色谱柱,以含0.01 mol/L乙酸铵的0.2%甲酸溶液-甲醇为流动相进行梯度洗脱,流速为0.60 mL/min,柱温为40℃,进样量为5μL;以可加热的电喷雾离子源为离子源,以全扫描-选择离子监测模式进行正离子扫描。采用该法对10家企业生产的15批β受体拮抗剂类药物原料药/制剂中N-亚硝基类杂质含量进行测定,并采用Discovery Studio软件对待测杂质进行毒性预测和关注阈值估算。结果5种β受体拮抗剂类药物中,N-亚硝基普萘洛尔、N-亚硝基美托洛尔、N-亚硝基阿替洛尔、N-亚硝基艾司洛尔、N-亚硝基比索洛尔检测质量浓度的线性范围分别为1.01~503.38、1.02~508.38、0.97~483.63、1.11~554.27、1.05~523.92 ng/mL(r>0.999),定量限分别为1.04、0.25、0.05、0.55、1.05 ng/mL,检测限分别为0.52、0.08、0.02、0.17、0.52 ng/mL,精密度、重复性、加样回收率、稳定性、耐用性试验的RSD均小于7.5%(n=6或n=5)。15批样品中,除1批样品外,其余批次均检出了N-亚硝基普萘洛尔(1.07~8.91 ng/mg)、N-亚硝基美托洛尔(1.43~3.37 ng/mg)、N-亚硝基阿替洛尔(1.33 ng/mg)、N-亚硝基艾司洛尔(0.19 ng/mg)、N-亚硝基比索洛尔(1.27 ng/mg)。经预测,上述5种杂质有不同程度的生育毒性、致突变性、致癌性,关注阈值分别为1.0、0.4、4.3、0.2、46.7 ng/mg。结论所建方法简单快捷、灵敏度高、专属性强,估算的关注阈值明确,可用于多种β受体拮抗剂类药物中N-亚硝基类杂质的含量控制。OBJECTIVE To determine the contents of N-nitroso impurities in raw materials/formulations of propranolol,metoprolol,atenolol,esmolol and bisoprolol,and clarify the attention threshold.METHODS Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q/Orbitrap HRMS)was adopted.An ACE Excel 3 C18-AR column was used for the separation and a mixture of 0.2%formic acid solution with 0.01 mol/L ammonium acetate and methanol was employed as the mobile phase by gradient elution,at a flow rate of 0.60 mL/min.The column temperature was set at 40℃,and the sample size was 5μL.The heated electrospray ionization source was employed in the positive full mass spectra-selected ion monitoring mode.The contents of N-nitroso impurities in raw materials/formulations of 15 batches ofβ-blockers from 10 manufacturers were determined by this method.Discovery Studio software was applied to predict the toxicity of the impurities and estimate the attention threshold.RESULTS Among 5 kinds ofβ-blockers,the linear ranges of N-nitroso propranolol,N-nitroso metoprolol,N-nitroso atenolol,N-nitroso esmolol and N-nitroso bisoprolol were 1.01-503.38,1.02-508.38,0.97-483.63,1.11-554.27 and 1.05-523.92 ng/mL,respectively(r>0.999).The limits of quantitation were 1.04,0.25,0.05,0.55 and 1.05 ng/mL,and the limits of detection were 0.52,0.08,0.02,0.17 and 0.52 ng/mL,respectively.RSDs of precision,reproducibility,recovery,stability and durability tests were all lower than 7.5%(n=6 or n=5).Among the 15 batches of samples,except for 1 batch,N-nitroso propranolol(1.07-8.91 ng/mg),N-nitroso metoprolol(1.43-3.37 ng/mg),N-nitroso atenolol(1.33 ng/mg),N-nitroso esmolol(0.19 ng/mg)and N-nitroso bisoprolol(1.27 ng/mg)were detected in all other batches.According to predictions,the above 5 impurities had varying degrees of reproductive toxicity,mutagenicity and carcinogenicity,with attention thresholds of 1.0,0.4,4.3,0.2 and 46.7 ng/mg,respectively.CONCLUSIONS The established method is simple,rap

关 键 词：β受体拮抗剂类药物 N-亚硝基类杂质 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱技术 基因毒性 关注阈值 

分 类 号：R917[医药卫生—药物分析学]