UPLC-QTOF-MS联用技术检测硫酸庆大霉素注射液中的乙二胺四乙酸二钠

目的：建立超高效亲水性液相色谱与电喷雾四极杆飞行时间串联质谱联用法（UPLC-QTOF-MS）对硫酸庆大霉素注射液中的辅料乙二胺四乙酸二钠进行分析。方法：以三氯化铁作为络合剂，色谱柱为Waters Xbridge HILIC柱（4.6 mm×250 mm，5 μm），流动相为乙腈：0.02 mol·L^-1醋酸铵水溶液（pH=3.0）梯度洗脱，检测波长为270 nm，柱温30 ℃。ESI离子源，负离子检测模式，离子源温度120 ℃。结果：乙二胺四乙酸铁（EDTA-Fe）络合物峰能与相邻未知组分峰很好地分离，乙二胺四乙酸二钠在1.671~80.198 μg·mL^-1范围内线性关系良好（r=0.9999）。LC-MS中，供试品质谱行为与对照品完全一致。结论：本法解决了乙二胺四乙酸二钠极性大，无紫外吸收，且常规反相色谱柱填料无法保留的难题。优化得到的色谱条件可以同时应用于LC-MS定性分析和LC-DAD定量测定，为乙二胺四乙酸二钠的检测工作提供了一定的参考依据。

Objective: To develop a new method for determination of disodium edetate in gentamycin sulfate injection by ultra performance hydrophilic interaction liquid chromatography-electrospray ionization-quadrupole time of flight/mass spectrometry (UPLC-QTOF-MS). Methods: The complexing agent was trichloride ferric.The separation was performed on a Waters Xbridge HILIC column(4.6 mm×250 mm,5 μm)and the mobile phase consisted of acetonitrile and 0.02 mol·L^-1 ammonium acetate solution (pH=3.0) for gradient elution.The detection wavelength was set at 270 nm and the column oven was maintained at 30℃.The mass spectrometry detector was equipped with an ESI source with a temperature of 120 ℃,and set at negative ion monitoring model. Results: Disodium edetate was completely separated from the impurities.The calibration curve of disodium edetate was linear in the range of 1.671-80.198 μg·mL^-1 (r=0.9999).The characteristics of samples were consistent with the reference substance according to the high performance liquid chromatogram and the mass spectrogram. Conclusion: The disodium edetate has high polarity,no UV absorption,and is difficult to be retained in the conventional rever sed phase column.The method can solve the above problem.The optimized chromatographic conditions can be applied to the qualitative LC-DAD and quantitative LC-MS and provide a reliable reference for future disodium edetate detection.