Ozenoxacin is an antibiotic drug prescribed to treat various skin infections caused by various bacteria. Various chemical mechanisms such as stille coupling, Buchwald–Hartwig coupling, cyclization and saponification are involved during the process of synthesis of Ozenoxacin. In the process of synthesis, there is a possibility of the formation of related impurities and among them, some are genotoxic impurities. To date, in literature, there is no method reported for analysing Potential Genotoxic Impurities (PGIs) in Ozenoxacin and hence this study was initiated to develop an LCMS method for quantification of two genotoxic impurities of Ozenoxacin viz., nitroso impurity, ester impurity. The analytes were resolved on Alltima C18 column (150×4.6mm; 5 μm particle size) using 0.01 mM ammonium acetate at pH 4.8 and methanol in 80:20 (v/v) at 0.5 mL/min flow rate and 10 μ sample injection volume. The multiple reaction monitoring of the mass fragments confirms the parent ion at m/z of 364, 393 and 496 for Ozenoxacin, Nitroso and Ester impurity respectively with characteristic product ion at m/z 196. The method has a linearity range of 0.05 μg/mL to 1.0 μg/mL for three analytes with detection limits of 0.015, 0.011 and 0.015 μg/mL for Ester impurity, Ozenoxacin and Nitroso impurity respectively. The method was validated and produces acceptable results and can successfully separate the potential genotoxic impurities in spiked commercial samples. Based on the findings, it was concluded that this method can be practically useful for the identification and quantification of potential genotoxic impurities and may apply to the safe use of Ozenoxacin in clinical treatment.