Abstract:
The aim of the present study was to develop and validate HPLC method and UV Spectrophotometeric method for the determination of allopurinol. In RP- HPLC a mixture of 70% buffer solution (mono basic ammonium phosphate 0.05M with 30% acetonitrile and methanol1:1) (v/v) was used as a modified mobile phase mixture. The column was C8 (250×4.6mm). The flow rate was set to 1ml/mint. The method was validated regarding linearity, range, limit of detection, limit of quantitation, precision, and accuracy. The obtained results showed that the validated method have good linearity, accuracy, precision and selectivity and reducing the retention time of allopurinol from (14.26) minutes to (3.0) minutes. Analytical method development results indicated that the assay (98.43%) and the limit of detection was (0.051µg/ml), limit of quantitation was (0.156µg/ml) and assay exhibited a linear over the range of (10-50µg/ml). The modified method of chromatography gave a linear relationship at a wavelength of 250 nm, linear correlation coefficient (0.9994). In the UV validated method using the buffer solution (pH 4.5). The method validated according to international conference on harmonization guideline, linearity range, and limit of detection, limit of quantitation, precision, and accuracy. The modified method of UV gave a linear relationship at a wavelength of 250 nm, linear correlation coefficient (0.9991).The validated method showed results the indicated limit of detection was (0.052µg/ml), limit of quantitation was (0.158µg/ml) and assay of UV validated method (100.15%) Respectively results Were compared with the official method were conducted analyzes in two different days to determine relative standard deviation which did not exceed 2%, the developed methods in present research successfully in the quantitation analysis of commercial preparation tablets of different sources. The study summarized the results of the analysis are not different from those obtained by official method approved therefore developed method considered to be fit, the best because fast, inexpensive and more safe, which can be of use for routine analysis drug lab.
