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Method development consists of selecting the appropriate wavelength and choice of stationary and mobile phase. The following studies were conducted for this purpose. The spectrum of diluted solutions of piperacillin and tazobactam in mobile phase were recorded separately on UV spectrophotometer. The peak of maximum absorbance wavelength was observed. The spectra of the standard drug showed that a wavelength was found to be 226?nm. The proposed method was validated as per ICH guidelines. The parameters studied for validation were specificity, linearity, precision, accuracy, robustness, system suitability, Vandetanib supplier limit of detection and limit of quantification. Under the experimental conditions described above, linear calibration curves for both piperacillin and tazobactam were obtained with six concentration levels each. The linearity range of piperacillin and tazobactam were 50�C100?ppm. 20?��L of each concentration was injected in duplicate into the HPLC system. The response was read at 226?nm and the corresponding chromatograms were recorded. The regression value of the plots was computed by least square regression method. Peak area (A) and concentration (C) of each drug substance was subjected to regression analysis to calculate the correlation coefficients. The correlation coefficient (r2?=?0.999 for piperacillin and r2?=?0.998 for tazobactam) of regression was found almost equal to 1. Linearity results are presented in Table 1 and graph in Fig.?2. Precision of the method was performed as intraday and interday precision. A standard solution of piperacillin and tazobactam were injected six times and corresponding peak areas were recorded. www.selleckchem.com/products/Bosutinib.html Results of system precision studies were shown in Tables 2 and 3. The % of RSD was found to be less than 2. The values of percentage of RSD obtained in intraday and interday precision were 0.681, 0.531 and 1.28, 1.405 for piperacillin and tazobactam respectively. The values of % of RSD within a day and day to day variation (Oxalosuccinic acid solution. Percent recovery was calculated by comparing the area before and after the addition of the standard drug. The standard addition method was performed at 50%, 75% and 100% level. The percent recovery and % RSD were calculated and results are presented in Table 4. Satisfactory recoveries ranging from 99.72 to 99.83 were obtained by the proposed method. The robustness was determined by carrying out the assay during which mobile phase ratio and pH of the mobile phase was altered slightly. When the pH was at 4.0, percentage of RSD was found to be 0.108 for piperacillin and 0.042 for tazobactam. On slight variation mobile phase ratio of upto ��10%, the change in percentage of RSD was 0.06 for piperacillin and 0.136 for tazobactam. At 224?nm wavelength, the percentage RSD was 0.69 for piperacillin and 1.
