The aim of this work is to establish a novel, simple, precise and sensitive UV-AUC spectrophotometry method for estimation of Pyrazinamide from bulk and in-Pharmaceutical formulation and validate it according ICH guidelines. Pyrazinamide is used for the treatment of tuberculosis. Methods and Methodology: Pyrazinamide was estimated by four simple UV-Spectrophotometric methods using double beam UV-Spectrophotometer (UV-2450, Shimadzu). A stock standard solution was prepared by dissolving 10 mg of Pyrazinamide in to 100 mL volumetric flask then volume was made by methanol up to the mark, the obtaining concentration was 100µg/mL. After suitable dilutions, it scans in the UV-visible range 200-400 nm. For Method A and Method B zero-order spectrum and area under curve (AUC) recorded at 268 nm and 259.40-273.20 nm respectively. While for Method C and for Method D first-order derivative the amplitude and area under curve (AUC) recorded at 279nm and 277.20-282.60 nm respectively. For linearity study, series of dilutions were prepared from stock solutions. Results: In Method A, B, C and Method D, Pyrazinamide followed linearity in the concentration range of 2-12 μg/mL with (r 2 > 0.999). The % recovery was found to be in the range of 98-102%. Furthermore, the precision of methods were calculated in terms of % RSD less than 2 showed, methods are precise. The developed methods was validate according to ICH guidelines. Conclusion: The developed methods are simple, rugged, robust and economical. The illustrated methods can routinely be used for determination of Pyrazinamide in bulk and in Pharmaceutical formulation.
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