Determination and Comparative Analysis of Aldehyde Genotoxic Impurity in Midazolam Active Pharmaceutical Ingredients and Midazolam Injections
Objective To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the content determination of 1-[4-chloro-2-(2-fluorobenzoyl)phenyl]-2-methyl-1H-imidazole-5-carbaldehyde(aldehyde-based genotoxic impurity M,referred to as impurity M)in midazolam active pharmaceutical ingredients(API)and midazolam injections,and to compare the content of impurity M in midazolam API and midazolam injections from three Chinese manufacturers,and reference preparations.Methods The chromatographic column was Acquity UPLC BEH-C18 column(100 mm × 2.1 mm,1.7 μm),the mobile phase was 0.01 mol/L aqueous ammonium formate-acetonitrile containing 0.1％formic acid(gradient elution),the flow rate was 0.4 mL/min,the column temperature was 40 ℃,and the injection volume was 5 μL.The electrospray ion source(ESI)was adopted with positive ion detection and multiple reactionmonitoring(MRM)mode,the capillary voltage was 3.5 kV,the temperature of desolvent was 350 ℃,the gas flow of the cone hole was 60 L/h,and the flow of the desolvent was 600 L/h.Results The linear ranges of impurity M were 0.46-9.16 ng/mL(r=0.999 2,n=7)and 28.62-1 144.77 ng/mL(r=0.995 3,n=7).The limit of detection was 0.03 ng/mL,and the limit of quantification was 0.13 ng/mL.The average recovery of impurity M was 98.53％with an RSD of 4.53％(n=6).The contents of impurity M in the Midazolam Injection from three domestic manufacturers were significantly higher than those in the midazolam API(P＜0.05),and the content of impurity M in the Midazolam Injection produced by manufacturer A,which did not pass the consistency evaluation,was the highest and significantly higher than that in the reference preparation(P＜0.05).Conclusion This method has high sensitivity and good repeatability,which can be used for quality control of impurity M in Midazolam Injection.It is recommended that manufacturers that have not passed consistency evaluation conduct consistency evaluation research as soon as possible,optimize preparation production processes,and formulate impurity control strategies based on risk assessment.
万方数据
维普
