Analytical Method Development and Validation for Determination of Assay of Antibacterial Drugs Besifloxacin Hydrochloride and Phenoxyethanol in Gel Formulation

Abstract

The present work aims to develop an analytical method and validate it to determine the assay of an antibacterial dug-in gel formulation. Background: Analytical Method Validation is a process involving confirmation studies that procedure/ method/ system/ analyst provides precise and reproducible outcome recognized by research laboratory studies that the performance features of the technique follows the necessities required for the analytical applications. Analytical Method Validation is a process involving confirmation studies that procedure/ method/ system/ analyst provides precise and reproducible outcomes recognized by research laboratory studies that the performance features of the technique follow the necessities required for the analytical applications. Analytical Method Validation is a process involving confirmation studies that procedure/ method/ system/ analyst provides precise and reproducible outcomes recognized by research laboratory studies that the performance features of the technique follow the necessities required for the analytical applications. Objective: To improve the conditions and parameters which should be followed in the development and validation by developing a new sensitive and accurate RP-HPLC method. Validating the proposed newly developed methods per the analytical parameters mentioned in the IP, USP, BP and ICH guidelines. HPLC method was validated to indicate that the analytical procedure used is suitable for intended use by using various parameters like specificity, linearity, LOD, LOQ, precision, accuracy, range, robustness, stability in analytical solution and system suitability. HPLC method was validated to indicate that the analytical procedure used is suitable for intended use by using various parameters like specificity, linearity, LOD, LOQ, precision, accuracy, range, robustness, stability in analytical solution and system suitability. The standard retention times for the Drug Besifloxacin were 7.781 min, and the sample was 7.731, respectively. The area of standard besifloxacin was 1828547, and the sample area was 1825315. The assay of the sample was 98%. The retention times for the drug Phenoxyethanol standard were found to be 2.010 min, and the sample was at 2.004, respectively. The Area of standard Phenoxyethanol was 438025, and the sample area was 438103. The assay of the sample was 97.04%. The RSD for 5 replicate injections for each peak is 0.33% in system suitability. In specificity, peaks of Diluent, Placebo & Impurities are not interfering with the Besifloxacin peaks. Peaks of Besifloxacin were found to be pure. Degradation products were found to be well separated from the besifloxacin peak. The peak purity factor was NLT 0.9995. In the precision study, the System Precision RSD of the Retention time for Besifloxacin obtained from six replicate injections was 0.33%. The RSD of the Area of Besifloxacin obtained from six replicate injections is 0.46%. Method precision RSD was calculated on 6 determinations assay value of Drug besifloxacin is 0.56%. The RSD calculated on 6 determinations for the assay value of the Drug besifloxacin is 0.50%. In Intermediate precision, RSD was calculated on 6 determinations for the assay value of the Drug besifloxacin is 0.50%. The RSD calculated on 12 determinations (Method precision & Intermediate precision) for assay value is 0.50%. Stability in the analytical solution for the standard and sample, the area difference of besifloxacin peak was within ±2.0% from initial Linearity. The correlation coefficient & regression coefficient (R square) should be not less than 0.995 for Besifloxacin Correlation Coefficient is 0.998 Regression coefficient is 1.000. The % intercept should be within ±5.0% of the response at 100% level Precision at 50% & 150% levels; the RSD is 0.01%. Precision at 50% & 200% level: the RSD was found to be NMT 2.0%. Accuracy means % recovery at each level found to be between 98 to 101 % of the drug besifloxacin. RSD on 9 (3 levels X 3) determinations is 1.2, following the NMT 2% range criteria. A correlation was 1.0% for the accuracy and linearity parameters. The % recovery is between 98% to 101%, and the % RSD for all recovery values is 1.41% which is within limits. The HPLC method optimized the conditions to obtain an adequate separation of eluted compounds. The % recovery is between 98% to 101%, and the % RSD for all recovery values is 1.41% which is within limits. The HPLC method optimized the conditions to obtain an adequate separation of eluted compounds.