Identification of Impurities in Thermally Degraded Cefmetazole Sodium for Injection with Two-Dimensional High-Performance Liquid Chromatography with Tandem Quadrupole Time-of-Flight Mass Spectrometry
ZOU Wen-bo1, CHEN Zhen-he2, WANG Ming-juan3*, DONG Jing2, ZHAO Xiao-jun3, JI Feng2, LI Xiao-dong2*, LIU Yu-jing3, CAO Lei2
1. National Institutes for Food and Drug Control, Beijing 102629, China; 2. Shimadzu Scientific Instrument China Innovation Center, Beijing 100020, China; 3. Beijing Sun-Novo Pharmaceutical Research Co.,Ltd., Beijing 102200, China
Abstract：OBJECTIVE To develop a two-dimensional high-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (2D-LC-QTOF MS) to identify impurities in thermally degraded cefmetazole sodium for injection. METHODS For the 1st dimensional chromatography, the mobile phase was a mixture of ammonium dihydrogen phosphate/tetrahydrofuran/methanol (730∶12.5∶300). Ammonium dihydrogen phosphate solution was prepared by dissolving 5.75 g salts in 730 mL water, then 19.2 mL tetrabutylammonium hydroxide (10%) was added in the above-mentioned solution. Finally, the solution was adjusted to pH 4.5 with phosphoric acid. The column used was Shim-pack GIS C18 (4.6 mm×250 mm, 5 μm), maintained at 25 ℃. The flow rate was 1 mL·min-1. For the 2nddimensional chromatography, the mobile phase was 1% formic acid aqueous solution (A)/acetonitrile (B), and gradient elution was conducted at a flow rate of 1.0 mL·min-1. The column was Shim-pack GIS C18(4.6 mm×150 mm, 5 μm) and installed in a 25 ℃ column oven. 50 μL quantitative ring was used to collect impurities from the 1st dimension. ESI source was used for ionization with both full scan and data dependent analysis mode (DDA). The collision gas was argon. Based on the accurate molecular weight of impurities and their products, isotope distribution, and fragmentation pathway for cefmetazole, the most reasonable molecular structures were proposed for unknown impurities. RESULTS Three unknown impurities were identified.Impurities 1 and 3 were isomers of cefmetazole, which were thermally degraded impurities of cefmetazole,meanwhile impurity 2 was 7-methylthiocefmetazole, which was from the production process. CONCLUSION The unknown impurities above identification threshold of ICH in cefmetazole sodium for injection were analyzed with 2D-LC-QTOF system, which was also suitable for the identification of unknown impurities in other β-lactam antibiotics using mobile phase containing non-volatile salts.
邹文博, 陈振贺, 王明娟, 董静, 赵小君, 冀峰, 李晓东, 刘宇晶, 曹磊. 采用二维高效色谱-串联四级杆飞行时间质谱法对注射用头孢美唑钠的未知杂质进行结构解析[J]. 中国药学杂志, 2022, 57(8): 645-650.
ZOU Wen-bo, CHEN Zhen-he, WANG Ming-juan, DONG Jing, ZHAO Xiao-jun, JI Feng, LI Xiao-dong, LIU Yu-jing, CAO Lei. Identification of Impurities in Thermally Degraded Cefmetazole Sodium for Injection with Two-Dimensional High-Performance Liquid Chromatography with Tandem Quadrupole Time-of-Flight Mass Spectrometry. Chinese Pharmaceutical Journal, 2022, 57(8): 645-650.
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2022, Vol. 57 
