核磁共振和液相色谱-质谱对氯唑沙宗未知杂质的结构分析及其检查方法研究  

Determination and structural analysis of unknown impurities in chlorzoxazone by NMR spectroscopy and liquid chromatography-mass spectrometry

作　　者：陈航 邓丽明[2] CHEN Hang;DENG Li-ming(Jiangsu Institute for Food and Drug Control,Nanjing 210019,China;Department of Pharmacy,Zhongda Hospital Affiliated to Nanjing Southeast University,Nanjing 210009,China)

机构地区：[1]江苏省食品药品监督检验研究院,南京210019 [2]东南大学附属中大医院药学部,南京210019

出　　处：《药物分析杂志》2021年第12期2132-2142,共11页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:采用核磁共振法和液相色谱-质谱对氯唑沙宗5个未知杂质进行结构确证,并建立有关物质检查的方法。方法:采用制备色谱分离提纯获得高纯度的杂质E,运用HPLC-MS、^(1)H-NMR、^(13)C-NMR、HSQC和HMBC等技术手段对5个杂质进行结构确证。有关物质检查采用Waters XBridge C_(18)柱(250 mm×4.6 mm,5μm),流动相为10 mmol·L^(-1)醋酸铵溶液-乙腈,梯度洗脱,流速为1.0 mL·min^(-1),检测波长为220 nm,柱温为30℃。结果:通过光谱数据分析,鉴定出氯唑沙宗原料药中5个杂质的结构,分别为苯并噁唑-2-酮(杂质A)、7-氨基-5-氯-苯并噁唑-2-酮(杂质B)、2-氨基-5-氯苯酚(杂质C)、5-溴-苯并噁唑-2-酮(杂质D)、7,7’-氯唑沙宗二聚体(杂质E)。结论:该研究为氯唑沙宗的质量控制提供了有效的方法依据。Objective:To identify and characterize the structures of five unknown impurities in chlorzoxazone by NMR and HPLC-MS,and establish a method to determine the related substances in chlorzoxazone.Methods:Impurity E was obtained by preparative HPLC,and the structures of five impurities were elucidated and confirmed by spectral data(HPLC-MS,^(1)H-NMR,^(13)C-NMR,HSQC and HMBC)comprehensively.The determination of related substances was performed at 30℃on a Waters XBridge C_(18)column(250 mm×4.6 mm,5μm)with 10 mmol·L^(-1)ammonium acetate solution-acetonitrile(gradient elution)was used as the mobile phase.The flow rate was 1.0 mL·min^(-1),and the detecting wavelength was 220 nm.Results:Based on the spectral data,the structures of five impurities were characterized as 2-benzoxazolinone(impurity A),7-amino-5-chloro-2-benzoxazolinone(impurity B),2-amino-5-chlorophenol(impurity C),5-bromo-2-benzoxazolinone(impurity D)and 7,7’-chlorzoxazone dimer(impurity E).An effective method is established to determine the related substances of chlorzoxazone.Conclusion:This established method is suitable for the identification of related substances and provides technical basis for the quality control of chlorzoxazone.

关 键 词：氯唑沙宗 核磁共振 高效液相色谱-串联质谱 杂质结构确证 制备色谱 有关物质 质量控制 

分 类 号：R917[医药卫生—药物分析学]