Development and validation of HPLC method for quantification of Cefotaxime in plasma of Patanwadi sheep

Present research work was carried out to study the validation procedure of Cefotaxime using plasma of Patanwadi sheep. The samples were analyzed using RP- HPLC C18 column (250 X 4.6 mm) with UV detection (254 nm). The method was developed for extraction of Cefotaxime from plasma of sheep using acetonitrile and was validated. The mobile phase was a mixture of 0.05 M potassium dihydrogen phosphate buffer with pH 6 and acetonitrile (88: 12, v/v). The method was validated with respect to linearity, precision and accuracy. The intra day and inter day precision study of cefotaxime was carried out by estimating the corresponding responses 3 times on the same day and on 3 different days (1st, 2nd and 3rd ) for 6 different concentration of Cefotaxime (10, 5, 2.5, 0.625, 0.312 and 0.156 ?g/ml) and the results were reported in terms of relative standard deviation (RSD). At all concentration studied the C.V. (Coefficient of Variance) was less than 7%. The recovery of Cefotaxime from plasma varied from 85-89%. The limit of detection (LOD) was found to be 0.156 ppm. Calibration graphs showed a linear correlation (r > 0.998) over the concentration ranges of 1.56 ? 200 ?g mL-1 for plasma. The results obtained indicated good precision of assay. The developed method was found to be simple, sensitive, accurate, precise and reproducible and can be used for analytical studies of Cefotaxime.