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Abstract

METHOD DEVELOPMENT AND VALIDATION OF AN ACTIVE METABOLITE OF PYRAZINAMIDE USING ULTRA VIOLET SPECTROPHOTOMETRY

Santosh Kumar Kushwaha*, Vivek Chauhan, Shikha Agnihotri, Pankaj Patel, Saurabh Shukla

ABSTRACT

The ultra violet (UV) spectrophotometry measures a particular wavelength absorbed by the sample from the UV spectral region. This method of analysis is very rapid and simple, however, lacks sensitivity and precision while analyzing samples of low concentrations. Despite lack of sensitivity of this method it is very useful when drug concentrations to be estimated are ample and need rapid detection. This method will be useful to estimate released drug from various solid dosage forms containing higher drug concentrations. Here in, we aim to develop a rapid method of analyzing an active metabolite of pyrazinamide viz., pyrazinoic acid (POA) using UV spectrophotometry. 4 μg/ml solution of POA in deionized water was scanned in the spectral range between 200-400 nm and the maximum absorbance was observed at wavelength of 268 nm. Samples of different concentrations analyzed for determining linearity demonstrated a linear response for a quantitation in the range of 2μg/ml to 10μg/ml in the deionized water. A regression equation; y=0.0585+0.019 and r2 =0.9997 was obtained after plotting the concentration vs. absorbance calibration curve. The method was validated for inter-day and intra-day accuracy, precision, detection limit, robustness and ruggedness. Furthermore, the method validated according to the ICH [Q2 (R1)] guidelines was found to be in acceptance limit. This quantification method is suitable for the estimation or regular quality check of POA in the bulk formulation like tablets, pharmaceutical powders and granules.

Keywords: UV-Spectrophotometry; Pyrazinoic acid; Method development; Validation; Accuracy.


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