Method validation The validation of analytical method verifies that the characteristics of the method if they satisfy the requirements of the method
The validation of analytical method verifies that the characteristics of the method if they satisfy the requirements of the method. The proposed method was validated according to ICH guidelines for specificity, linearity, accuracy, and precision, and robustness. Specificity was carried out in which no interference of the excipients was observed at retention time of the analytical peak. Calibration curve was constructed by plotting concentration Vs plot area. It showed that there was a good linear relationship in the concentration range of 40% to 160% with > 0.999 as the value of correlation co-efficient. The accuracy of the method was studied by analyzing the drug solutions at 80%, 100% and120% concentration level. The mean percentage recovery was found to be 101.7% For precision the sample solution at working concentration was analyzed in replicate as per the method. The percentage relative standard deviation was found to be less than 1%. Robustness of the method shows no significant change in system suitability parameters and mean % assay at modifying chromatographic conditions from the original method
The Specificity of the method was established by injecting a Blank, (diluent) Placebo and Sample Preparations into the chromatograph. No interference was observed. This indicates that the solvent does not interfere with drug peak and shows a good resolution.
The peak area responses of all solutions over concentration levels ranging from 40% to 160% of target concentration were measured in triplicate. Linear correlation was obtained between peak area and concentration of Erythromycin estolate for LC method. The linearity of the calibration curves was validated by the value of correlation coefficients of the regression
(r = 0.9999).The regression analysis of the calibration curves is shown in Figure 3.