Abstract

Anita Patidar, Dr. Priydarshani R. Kamble*

ABSTRACT

A broad-spectrum aminoglycoside antibiotic, amikacin is often used to treat severe Gram-negative infections and TB that has developed resistance to many drugs. An amikacin detection technique that is sensitive, selective, and robust was developed and validated in this work using liquid chromatography-mass spectrometry (LC-MS). A reversed-phase C18 column was used for chromatographic separation under isocratic conditions with a mobile phase consisting of acetonitrile and 0.1% formic acid in water. A 10 µL injection volume resulted in an optimal flow rate of 0.8 mL/min. The electrospray ionization was used in positive mode for detection, with the transition from precursor to product ions of amikacin (m/z 586 ? 163) being monitored in multiple reaction monitoring (MRM). There was zero interference from excipients or endogenous drug sample components during the retention time of amikacin, demonstrating good specificity for the approach. Correlation coefficients (R²) regularly exceeded 0.999, and calibration curves were linear in the concentration range of 0.05-50 µg/mL. The method's great sensitivity was confirmed by the determination of limits of detection (LOD) and quantification (LOQ) as 0.02 µg/mL and 0.05 µg/mL, respectively. tests on precision, both within and between days, showed RSDs below 2%, while tests on accuracy showed recoveries between 98% and 102% over a range of concentrations. Consistent results were obtained when the method's robustness was tested under controlled conditions, such as changing the mobile phase composition, column temperature, and flow rate. Amikacin was shown to be stable in processed samples for a minimum of 24 hours at room temperature and for a maximum of one month when frozen, according to stability tests. This validated LC-MS approach was used to quantify amikacin in commercial injectable formulations and spiked human medication samples, showing that it is useful for clinical pharmacokinetic research and pharmaceutical quality assurance. Amikacin bioequivalence research, therapeutic medication monitoring, and routine analysis may all benefit from the method's selectivity, repeatability, and sensitivity.