目的:以超高效液相色谱-四极杆/静电场轨道阱质谱(UPLC-Q-Orbitrap HRMS)技术为主要手色谱段,结构定性亚硫酸氢钠甲萘醌注射液中的4 个降解杂质。方法:超高效液相色谱采用Thermo Gold C18 色谱柱(100 mm×2.1 mm,1.9 μm),以0.02 mol·L-1 乙酸铵溶液-甲醇(92∶8)为流动相,质谱采用HESI 离子源、正负离子检测模式,碎片分析借助MsFrontier7.0 软件,结构定性分析亚硫酸氢钠甲萘醌注射液中的4 个降解杂质。结果:通过HPLC-PDA、UPLC-HRMS、1HNMR 等手段,结合有机合成制备杂质对照品、强制降解试验归属杂质来源、推测形成路径等方式,确定了亚硫酸氢钠甲萘醌的4 个重要的降解杂质的分子结构,它们分别是：3-羟甲基-1,4-二-氧代-1,4-二-氢-2-萘磺酸钠(杂质1)、1-羟基-3-甲基-4-氧代-1,2,3,4-四-氢萘-1,3-二-磺酸钠(杂质2)、4-羟基3-甲基-1-氧代-1,2-二-氢-萘磺酸钠(杂质3)和1,4-二-羟基3-甲基-2-萘磺酸钠(杂质4)。结论:通过降解杂质结构定性及其互变异构、自氧化等机制研究,为亚硫酸氢钠甲萘醌注射液的工艺优化和质量控制提供了参考依据。
Objective: To confirm the structure of four degradation impurities of menadione sodium bisulfite injection using UPLC-Q-Orbitrap HRMS. Methods: UPLC was applied with a Thermo Gold C18 column(100 mm×2.1 mm,1.9 μm) and eluted with 0.02 mol·L-1 ammonium acetate solution-methanol (92∶8). MS was applied with HESI source and positive/negative ion modes. Fragment analysis was carried out by Mas Frontier7.0 software. The structures of four degradation impurities in the injection of menadione sodium bisulfite were characterized and confirmed. Results: Four degradation impurities,which were sodium 3-(hydroxymethyl) -1,4-dioxo-1,4-dihydronaphthalene-2-sulfonate (impurity 1),sodium 1-hydroxy-3-methyl-4-oxo-1, 2,3,4-tetrahydronaphthalene-1,3-disulfonate (impurity 2),sodium 4-hydroxy-3-methyl-1-oxo-1, 2-dihydronaphthalene-2-sulfonate (impurity 3) and sodium 1,4-dihydroxy-3-methylnaphthalene-2-sulfonate (impurity 4), were confirmed. This study used analytical methods such as HPLC-PDA,UPLC-HRMS,and 1HNMR. The degradation mechanism was proposed and confirmed by organic synthesis,forced degradation etc. Conclusion: Through the structure study of the degradation impurities and the mechanism of tautomerism and auto-oxidation,this method provides a reference for the process optimization and quality control of sodium bisulfite menadione injection.
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