超高效液相色谱-四极杆静电场高分辨质谱联用法测定盐酸普萘洛尔缓释片中痕量N-亚硝基普萘洛尔  

Determination of Trace N-nitrosopropranolol in Propranolol Hydrochloride Sustained Release Tablets by UHPLC-Q-Orbitrap HRMS

作　　者：郭常川 文松松 吕邓义 王维剑 杨书娟 牛冲 徐玉文 GUO Changchuan;WEN Songsong;LYU Dengyi;WANG Weijian;YANG Shujuan;NIU Chong;XU Yuwen(Shandong Institute for Food and Drug Control,National Medical Products Administration(NMPA)Key Laboratory for Research and Evaluation of Generic Drugs,Shandong Research Center of Engineering and Technology for Consistency Evaluation of Generic Drugs,Shandong Engineering Research Center of Transdermal Drug Delivery System,Industrial Technology Foundation Public Service Platform,Jinan 250101,China;Shandong Jewim Pharmaceutical Co.,Ltd,Taian 271000,China)

机构地区：[1]山东省食品药品检验研究院、国家药品监督管理局仿制药研究与评价重点实验室、山东省仿制药一致性评价工程技术研究中心、经皮给药系统山东省工程研究中心、产业技术基础公共服务平台,济南250101 [2]山东京卫制药有限公司,泰安271000

出　　处：《医药导报》2025年第4期628-633,共6页Herald of Medicine

基　　金：国家自然科学基金资助项目(81573606,81973486);泰山产业领军人才工程专项经费(tscx202306073)。

摘　　要：目的建立测定盐酸普萘洛尔缓释片中遗传毒性杂质N-亚硝基普萘洛尔(NPPN)的超高效液相色谱-四极杆静电场高分辨质谱联用检测方法(UHPLC-Q-Orbitrap HRMS)。方法供试品以甲醇作为溶剂超声提取,经离心和滤过后进样分析。采用2.7μm细粒径C 18超高效柱进行色谱分离,流动相为0.1%甲酸水和0.1%甲酸乙腈,程序洗脱。质谱采用HESI离子源,正离子PRM扫描模式,监测NPPN子离子m/z 72.0808,标准曲线法定量。结果NPPN在0.51~20.30 ng·mL^(-1)浓度范围内线性关系良好,相关系数(r)为0.9999;回收率为95.4%~98.3%,RSD为2.5%~4.2%,检出限0.20 ng·mL^(-1),定量限0.51 ng·mL^(-1)。应用该方法对6批盐酸普萘洛尔缓释片中的NPPN进行测定,6批样品中均检出了NPPN,其中3批检出量超过了可接受限度。结论该方法灵敏、准确、快速,可帮助药企进行生产工艺控制,并为药监部门提供有力的技术支撑。Objective To establish a ultrahigh-performance liquid chromatography-orbitrap high-resolution mass spectrometry(UHPLC-Orbitrap HRMS)method for the determination of the genotoxic impurity N-nitroso propranolol(NPPN)in propranolol hydrochloride sustained-release tablets.Methods The test sample was ultrasonically extracted using methanol as the solvent,then centrifuged and filtered before injection analysis.Chromatographic separation was performed using a 2.7μm particle size C 18 UHPLC column with a mobile phase of 0.1%formic acid(A)in water and 0.1%formic acid(B)in acetonitrile,using gradient elution.Mass spectrometry was conducted with an HESI ion source in positive ion parallel reaction monitoring(PRM)scan mode,monitoring the NPPN fragment ion at m/z 72.0808,and quantification was performed using the standard curve method.Results The calibration curve was in good linearity in the range of 0.51-20.30 ng·mL^(-1) with excellent correlation coefficient(r)of 0.9999.The recoveries of NPPN at three levels(low,medium,and high)were in the range of 95.4%~98.3%,while the RSDs were from 2.5%to 4.2%.The limit of detection(LOD)was 0.20 ng·mL^(-1) while the limit of quantitfication(LOQ)was 0.51 ng·mL^(-1).This analytical method was used to determine NPPN in six batches of propranolol hydrochloride sustained release tablet samples.NPPN was detected in all six samples,among which the detection amount of 3 batches have exceeded the acceptable limit.Conclusion This method is sensitive,accurate,and fast,making it useful for pharmaceutical companies in controlling production processes and providing robust technical support for regulatory authorities.

关 键 词：N-亚硝基普萘洛尔 遗传毒性 杂质 高分辨质谱 超高效液相色谱 含量测定 

分 类 号：R917[医药卫生—药物分析学] R927.2[医药卫生—药学]