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Objective: Development and validation of four simple, accurate, selective and sensitive UV spectrophotometric methodsfor the determination of rafoxanide in the presence of its alkali-induced degradation product without preliminary separation. Methods: (A) Dual wavelength method, where the difference in absorbance were calculated at 242 and 281 nm. (B) Area under the curve method,where area under the peak were measured at wavelength ranges (225-235 nm) and (275-295 nm). (C) Bivariate method, where the absorbance values were measured at selected wavelengths 250 and 280 nm. (D) First derivative method, whereThe amplitudes were measured at 253 nm. Results: Good linearity was found in the concentration range of 2.5–25 μg/mL for all the methods. The specificity of the methods were assessed by analysing synthetic mixtures containing the drug and its degradation product. Methods are validated as per ICH guidelines and accuracy, precision, repeatability and robustness are found to be within the acceptable limit. Conclusion: The proposed methods have been found to be accurate, precise and can be used for determination of the drug in pure form and pharmaceutical formulations as well as in the presence of its degradation product without any preliminary separation steps.

Quantitative Estimation and Validation of Rafoxanide in Presence of its Alkali-Induced Degradation Product by Different Spectrophotometric Methods