Background: Amikacin sulfate (AMK) belongs to the class of aminoglycoside antibiotics. It
is effective against the infections caused by Gram-negative and positive bacteria. AMK lacks a chromophore
group in its structure and, therefore, it does not absorb light in the 200-800 nm region which
makes it a difficult molecule to analyze by UV detector using high performance liquid chromatography
Objective: This study has been carried out to develop and validate a relatively simple, accurate, precise,
rapid, economical, and stability-indicating pre-column derivatization HPLC method for the determination
of AMK in pure and parenteral dosage forms.
Method: The stock solution of AMK was derivatized prior to its analysis. The mobile phase used for the
analysis was acetonitrile and water in the ratio of 50:50 (v/v) at pH 6.0. The method has been validated
according to the guideline of International Council for Harmonization (ICH) and different parameters
such as linearity, range, accuracy, precision, sensitivity, robustness, solution stability, specificity and
system suitability have been studied. AMK was subjected to stress degradation studies including thermolysis,
humidity exposure, acid-base hydrolysis, and oxidation in order to determine the specificity of
the test method.
Results: The retention time of AMK has been found to be 4.7 min. The results indicated that the method
is linear in the concentration range of 12.5-125% and possesses high accuracy (99.88±0.42%), precision
(<1.2%) and robustness (<0.5%). The obtained results are compared statistically with a reference
Conclusion: It was observed that the stress degradation studies do not affect the accuracy of the
method. Hence the proposed method can be used for the assay of AMK and its parenteral dosage form.