Abstract: | Abstract Objective To establish a pre-column derivatization HPLC method for the determination ofhydrazine in amikacin sulfate and its preparations. Methods 5% benzaldehyde methanol solution was used as thederivatization reagent. Agilent Zobax Eclipse XDB-C18 column (2.1 mm×150 mm, 5 μm) was used. The mobilephase A was 0.1% formic acid solution, the mobile phase B was acetonitrile, and linear gradient elution was carriedout according to the attached table. The flow rate was 1.0 mL/min. DAD UV detector was used, and the detectionwavelength was 300 nm. The column temperature was 25℃, and the injection volume was 20 μL. Results Thelinear relationship of hydrazine was found in the concentration range of 0.5022~12.5543 μg/mL(r=0.9994). Therecovery values of amikacin sulfate, amikacin sulfate for injection, and amikacin sulfate injection were 94.8%, 94.1%,and 94.3%, respectively. The content of hydrazine in amikacin sulfate, amikacin sulfate for injection, and amikacinsulfate injection in domestic enterprises were undetected~1.8 μg/g, undetected~0.2 μg/g, and undetected~3.1 μg/g,respectively. No hydrazine was detected in amikacin sulfate for injection and amikacin sulfate injection in nichiikopharmaceutical Co. Ltd. Conclusion The new precolumn derivatization HPLC method was accurate, simple, andspecific. It can be used to control the genotoxic impurity hydrazine in amikacin sulfate and its |