Purpose: To validate a UV-visible spectrophotometric technique for evaluating niclosamide (NIC) concentration in different media across various values of pH. Methods: NIC was investigated using a UV-visible spectrophotometer in acidic buffer solution (ABS) of pH 1.2, deionized water (DW), and phosphate buffer solution (PBS), pH 7.4. The characterization of NIC was done with differential scanning calorimeter (DSC), powder X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The UV analysis was validated for accuracy, precision, linearity, and robustness. Results: The DSC spectra showed a single endothermic peak at 228.43 °C (corresponding to the melting point of NIC), while XRD and FTIR analysis confirmed the identity, crystallinity and purity of NIC. In all media, the measured concentration of NIC was within ± 5 % of the actual value, which confirmed accuracy. The percentage relative standard deviation values were < 1 %, reflecting the precision of the method. The range of concentration measured was between 2 and 24 μg/mL, and all coefficient of determination (R2) values were > 0.99, indicating the linearity of the established analytical method. The limit of detection (LOD) and limit of quantification (LOQ) values were 0.122 and 0.407 μg/mL in ethanol, 0.530 and 1.766 μg/mL in ABS (pH 1.2), 0.224 and 0.747 μg/mL in DW, and 0.798 and 2.662 μg/mL in PBS, pH 7.4. The robustness was confirmed as the measured concentration under slight changes in temperatures and wavelengths were insignificant (p > 0.05). Conclusion: Based on the results above, the UV-visible spectrophotometric method under investigation was validated to be accurate, precise, linear, and robust in all the different media for the determination of NIC.
