Background A simple, rapid, accurate, and sensitive spectrometric method for the determination of Marbofloxacin has been developed. The present manuscript describes a first, second, third, and fourth derivative spectrophotometric method for the simultaneous estimation of and Marbofloxacin. Results First order derivative absorption at wavelengths of 300 nm and 278 nm, second order derivative absorption at wavelengths of 310 nm, 291 nm, and 270 nm, third order derivative absorption at wavelengths of 262 nm, 282 nm, 297 nm, and 311 nm, and fourth order derivative absorption at wavelengths of 258 nm, 267 nm, 292 nm, 304 (zero cross point for Marbofloxacin). With a correlation value (R2) of 0.994, marbofloxacin linearity was achieved in the concentration range of 2–20 µg/ml. The average recovery rate was found to be between 98.25 and 101.52%. Conclusions The suitability of these methods for the quantitative determination of Marbofloxacin was proved by validation. The proposed method was validated in accordance with the guidelines and was successfully applied to the simultaneous estimation of Marbofloxacin in its synthetic forms.

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