| Analytical Method Validation |
1.0 OBJECTIVE To lay down a procedure for Performance developing the analytical method validation. 2.0 SCOPE This SOP is applicable for developing the analytical method validation in Quality Control Department. 3.0 RESPONSIBILITY 3.1 QC – Officer/ Sr. Officer / Executive and above. 4.0 ACCOUNTABILITY Head-QC 5.0 PROCEDURE 5.1 Pre-Requisite: 5.1.1 All the instrument used in AMV must be calibrated. 5.1.2 Analyst qualification must be ensured who performing the AMV. 5.1.3 All the measuring glassware used in AMV must be calibrated or class ‘A’ type. 5.1.4 Analytical Method Validation is performed like Assay, Dissolution, impurities/related substances individually. 5.1.5 In case of Assay or impurity profile, system suitability is essential prior to start the validation activity. 5.1.6 Any analytical method shall be revised through change control procedure and shall be duly approved by Quality Assurance and then it undergoes for validation. 5.1.7 Whenever there is need to change critical test parameters, the method is re-validated with the revised parameters & conditions. 5.2 Specificity:(Analytical Method Validation) 5.2.1 Specificity is the ability to assess unequivocally the analytic, in the presence of components that may be expected to be present, such as impurities, degradation products and matrix components. 5.2.2 Specificity shall be determined by injecting blank, gradient blank, (If applicable), Standard solution, Sample, Individual known impurities. Perform peak purity check for analyte peak. (For HPLC with PDA Detector). 5.3 Forced Degradation: 5.3.1 Forced degradation is a degradation of new drug product at conditions more severe than accelerated conditions. It is required to demonstrate specificity of stability sample indicating methods & also provide an insight into degradation pathways and degradation products of the drug substance and helps in elucidation of the structure of the degradation products. Forced degradation studies show the chemical behaviour of the molecules, which in turn helped in the development of formulation and package. In addition, the regulatory guidance is very general and does not explain about the performance of forced degradation studies in QC. Thus, this review discusses the current trends in performance of forced degradation studies by providing a strategy for conducting studies on degradation mechanisms and describes the analytical methods helpful for development of stability indicating method. • Acid degradation • Alkali degradation • Peroxide degradation • Thermal • degradation • UV degradation • Humidity degradation 5.4 Precision: 5.4.1 Precision is the measure of how close the data values to each other for a number of measurements under the same analytical conditions. • System Precision • Method Precision • Intermediate Precision (Ruggedness) • FilterEquivalency 5.4.2 System Precision shall be determined by injecting six replicates of standard preparation as per method % RSD shall be calculated for peak area. 5.4.3 Method precision shall be performed by injecting six test solution preparations as per the methodology representing a single batch Determine the results % RSD for solvent content shall be calculated. 5.4.4 Intermediate Precision (Ruggedness) is the precision obtained when the assay is performed by multiple analyst, using multiple instruments or multiple days in one laboratory. Different sources of reagents and multiple lots of columns are included in this study. Intermediate Precision results are used to identify which of the above factors contribute significant variability to the final result in quality control lab. Depending upon time and resources, the method can be tested on multiple day’s analyst instrument etc. 5.5 Accuracy: 5.5.1 Accuracy is the exactness of an analytical method or the closeness of agreement between the value (which is accepted either as a conventional true value or an accepted reference and value found. It are measured as the percent of analytic recovered from matrix components). 5.5.2 For the assay of the drug substance, accuracy measurements are obtained by comparison of the results with the analysis of a standard reference material or by comparison to a second well-characterized method. 5.5.3 For the assay of the drug product, accuracy is evaluated by analyzing synthetic mixtures spiked with known quantities of components. 5.5.4 For the quantization of impurities, accuracy is determined by analyzing sample (drug substance or drug product) spiked with known amounts of impurities. 5.6 Linearity and Range: 5.6.1 To demonstrate that the analytical method is capable to obtain test results, which are directly proportional to the concentration of analytic. 5.6.2 Provide acceptable degree of linearity, accuracy and precision when applied to samples containing amounts of analytic within or at the extreme of the specified range of the analytical procedure. 5.6.3 Linearity shall be determined at minimum five concentration ranges from 50 % to 150 % of the specification limit. Linearity graph shall be plotted between responses versus concentrations. Squared correlation coefficient (Correlation coefficient, Residual sum of square, slope and Y-intercept) shall be calculated. 5.7 Robustness: 5.7.1 To demonstrate that the analytical method is capable to yield reproducible results under small but deliberate variations in method parameters such as column temperature, flow rate and buffer concentration. 5.8 Stability of solution: 5.8.1 The period of time a solution can be held before analysis without compromising accuracy. This delay is beyond that included in the method procedure anticipates unexpected instrumental delays. 5.8.2 The solution stability shall be determined by analyzing standard, test solution at specification level for initial stage and at various periodic intervals up to about 24 hours. Area response of initial sample (zero hours) shall be compared with periodic interval area response. The study may be stopped in the case of two consecutive criterion failures of standard, test solution. If failure observed at any single time interval only, then this shall not be considered as failure and sequence can be continued to next intervals 5.8.3 The temperature at which solution stability shall be performed and nature of glassware to be used shall be defined in the protocol. 5.8.4 The standard, test solution and spiked sample solution shall be considered stable for the time period till which the area difference between initial and next periodic interval RSD shall be not more than 2 % for Assay and 5% for Related Substance. |