HPLC-DAD-MS法对酮咯酸氨丁三醇及其注射液有关物质的分析

目的：对国内各厂家生产的酮咯酸氨丁三醇及注射液的有关物质现状进行考察，为标准提高提供依据。方法：建立液相色谱-二极管阵列检测器-质谱联用法，采用C₁₈（100 mm×4.6 mm，3 μm）色谱柱，以0.05 mol·L^-1乙酸铵溶液（甲酸调节pH至2.5）-乙腈（60：40）为流动相，对酮咯酸氨丁三醇及注射液中有关物质进行分离，采用二极管阵列检测器及ESI/Q/TOF高分辨质谱检测器进行检测，对检测到的有关物质进行结构解析，并将有关物质检测结果与现行标准方法检测结果进行对比。结果：在所建立的液质条件下，酮咯酸氨丁三醇及其有关物质分离良好，可以检测到酮咯酸氨丁三醇合成副产物及降解产物共22个，推断了有关物质1、4、6、10、13、15、21和22的结构。现有酮咯酸氨丁三醇原料药及注射液产品中实际存在的有关物质有13个。现行标准方法对酮咯酸氨丁三醇及注射液中的有关物质没有漏检。结论：液质联用技术能有效地分离鉴定酮咯酸氨丁三醇及注射液中的有关物质，液质分析结果为酮咯酸氨丁三醇及注射液的质量控制提供了参考。

Objective: To analyze the related substances in the domestic products of ketorolac tromethamine and ketorolac tromethamine injection for the purpose of standard improvement.Methods: An HPLC-DAD-MS method was developed. A C₁₈ column(100 mm×4.6 mm, 3 μm)was used for separation of the related substances with a mixture of acetonitrile and ammonium acetate buffer(containing 0.05 mol·L^-1 ammonium acetate, adjusting pH to 2.5 with formic acid)as the mobile phase. The related substances were detected by DAD and ESI/Q/TOF high resolution MS detector. The structure elucidation of the related substances was based on the results of MS and MS/MS detection. And the results were compared with the results acquired from the tests using HPLC methods in current standards.Results: Ketorolac tromethamine and its related substances were separated and detected under the established HPLC-DAD-MS condition. 22 related substances of ketorolac tromethamine were detected. 13 of them actually existed in the ketorolac tromethamine and ketorolac tromethamine injection products. The structures of related substance 1, 4, 6, 10, 13, 15, 21 and 22 were analyzed preliminarily. 5 of them were included in USP or BP and one of them was documented while 2 of them were newly discovered. All the related substances could be detected by HPLC methods in current standards.Conclusion: The HPLC-MS technology were proved to be effective in separation and identification of the related substances in ketorolac tromethamine and ketorolac tromethamine injection. The results of HPLC-DAD-MS were useful for quality control.