西洛他唑有关物质的色谱-质谱结构鉴定

目的:采用色谱-质谱联用技术对西洛他唑有关物质进行结构鉴定。方法:采用Waters XTerra RP-8(100 mm×4.6 mm,3.5μm)色谱柱串联Agilent Eclipse XDB-C₈(12.5 mm×4.6 mm,5μm)保护柱,以水-乙腈为流动相梯度洗脱,流速为1.0 mL·min^-1,检测波长为254 nm,对西洛他唑有关物质进行分离;采用电喷雾正离子化的方法,通过飞行时间质谱(TOF)测定一级质谱精确质量,三重四极杆/线性离子阱质谱(Q-TRAP)扫描二级与三级质谱,并进行结构解析。飞行时间质谱(TOF)和三重四极杆/线性离子阱质谱(Q-TRAP)的喷雾电压、雾化气压力、去溶剂温度依次分别为4 kV和5.5 kV、240 kPa和275 kPa、350℃和480℃。结果:西洛他唑与其有关物质分离良好,检测出8个有关物质,其中4个尚未见文献报道,并综合分析鉴定了有关物质结构。它们大部分具有6-[4-(1-环己基-1H-戊四唑-5-基)丁氧基-2(1H)-喹诺酮结构,制剂中有关物质状态与原料药中的相似。结论:色谱-质谱联用技术能有效地鉴定西洛他唑中有关物质,为其工艺和质量控制提供参考依据。

Objective:To identify the related substances in cilostazol by high performance liquid chromatography coupled to time-of-flight mass spectrometry(TOF) and triple quadrupole-linear ion trap mass spectrometry(Q-TRAP) hyphenated techniques.Methods:A Waters XTerra RP-8(100 mm×4.6 mm,3.5 μm)column and a Agilent Eclipse XDB-C₈(12.5 mm×4.6 mm,5 μm) guard column were used for the separation of the related substances. The mobile phase consisted of acetonitrile and water using a gradient program at a flow rate of 1.0 mL·min^-1 with 254 nm detection. Electrospray ionization source was operated in positive ion mode. The structures of the related substances were speculated by TOF for accurate ion mass determination and by Q-TRAP for product ion scanning. The conditions of TOF and Q-TRAP were as follows:the spray voltage 4 kV and 5.5 kV,the nitrogen draying gas nebulizer pressure 240 kPa and 275 kPa,the gas temperature 350℃ and 480℃,respectively.Results:Cilostazol and the related substances were well separated. Eight related substances in cilostazol,4 in which had not been reported previously,were characterized mostly as the same frame structure of 6-[4-(1-cyclohexyl-1H-tetrazol-5-yl)butoxy]-2(1H)-Quinolinone by hyphenated techniques. The impurity profiles for the preparations were similar with the API.Conclusion:The hyphenated LC-MS method is useful for the identification of related substances in cilostazol. The results obtained are valuable for its manufacturing process and quality control.

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