Potency Method Validation PDF

POTENCY METHOD VALIDATION MOKARAM HOSSAIN 30-Nov-24 1 Life cycle of an analytical method MOKARAM HOSSAIN 30-Nov-24 2 Guidelines for Drug Potency Assay MOKARAM HOSSAIN 30-Nov-24 3  Types of Quantitation  Quantitation by...

POTENCY METHOD VALIDATION MOKARAM HOSSAIN 30-Nov-24 1 Life cycle of an analytical method MOKARAM HOSSAIN 30-Nov-24 2 Guidelines for Drug Potency Assay MOKARAM HOSSAIN 30-Nov-24 3  Types of Quantitation  Quantitation by External Standard  Quantitation by Internal Standard MOKARAM HOSSAIN 30-Nov-24 4  Standard Plots for Quantitation  Single-Point Calibration  Multiple-Point Calibration  One Standard Calibration for Each Strength. MOKARAM HOSSAIN 30-Nov-24 5  System Suitability Requirements for Potency Assay  System suitability is a measure of the performance of a given system on a given day within a particular sample analysis set.  System suitability is part of method validation. MOKARAM HOSSAIN 30-Nov-24 6  System Suitability Requirements for Potency Assay  The main objective of system suitability is to recognize whether or not system operation is adequate given such variability as chromatographic columns, column aging, mobile-phase variations, and variations in instrumentation. MOKARAM HOSSAIN 30-Nov-24 7  System Suitability Requirements for Potency Assay  A system suitability test should be performed in full each time a system is used for an assay. If the system is in continuous use for the same analysis over an extended period, system suitability should be reevaluated at appropriate intervals to ensure that the system is still functioning adequately for its intended use. MOKARAM HOSSAIN 30-Nov-24 8  System Suitability Requirements for Potency Assay  System suitability should be based on criteria and parameters collected as a group that will be able to define the performance of the system. Some of the common parameters used include precision of repetitive injections (usually five or six), resolution (R), tailing factor (T ), number of theoretical plates (N), and capacity factor (k). MOKARAM HOSSAIN 30-Nov-24 9  Stability Indicating Potency Assay  content uniformity assay MOKARAM HOSSAIN 30-Nov-24 10  Linearity  Range  Accuracy  Precision *Repeatability (Precision) *Intermediate Precision *Reproducibility.  Robustness  Specificity MOKARAM HOSSAIN 30-Nov-24 11  Procedure  Linearity is usually demonstrated directly by dilution of a standard stock solution.  It is recommended that linearity be performed by serial dilution of a common stock solution.  Linearity is best evaluated by visual inspection of a plot of the signals as a function of analyte concentration. MOKARAM HOSSAIN 30-Nov-24 12  Subsequently, the variable data are generally used to calculate a regression by the least squares method.  the usual range for the potency assay of a drug substance or a drug product should be ±20% of the target or nominal concentration and ±30% for a content uniformity assay.  At least five concentration levels should be used. MOKARAM HOSSAIN 30-Nov-24 13  The slope, residual sum of squares, and y-intercept should also be reported as required by the ICH.  The slope of the regression line will provide an idea of the sensitivity of the regression and hence the method to be validated.  The y-intercept will provide the analyst with an estimate of the variability of the method. MOKARAM HOSSAIN 30-Nov-24 14 Acceptance Criteria  linearity is achieved when the coefficient of determination (r2) is ≥0.997 MOKARAM HOSSAIN 30-Nov-24 15 Linearity with correlation Linearity with correlation coefficient greater than 0.997 coefficient less than 0.997 MOKARAM HOSSAIN 30-Nov-24 16  Procedure  The specified range is normally derived from linearity studies and depends on the intended application of the procedure.  It is established by confirming that the analytical procedure provides an acceptable degree of linearity, accuracy and precision when applied to samples containing amounts of analyte within or at the extremes of the specified range of the analytical procedure. MOKARAM HOSSAIN 30-Nov-24 17  for the assay of a drug substance or a finished (drug) product: normally from 80 to 120 percent of the test concentration  for content uniformity, covering a minimum of 70 to 130 percent of the test concentration, unless a wider more appropriate range, based on the nature of the dosage form (e.g., metered dose inhalers), is justified MOKARAM HOSSAIN 30-Nov-24 18  Procedure  The ICH also recommended assessing a minimum of nine determinations over a minimum of three concentration levels covering the specified range (e.g., three concentrations/ three replicates)  For a drug substance, the common method of determining accuracy is to apply the analytical procedure to the drug substance and to quantitate it against a reference standard of known purity. MOKARAM HOSSAIN 30-Nov-24 19  For the drug product, accuracy is usually determined by application of the analytical procedure to synthetic mixtures of the drug product components or placebo dosage form to which known quantities of drug substance of known purity have been added.  The range for the accuracy limit should be within the linear range. MOKARAM HOSSAIN 30-Nov-24 20  Acceptance Criteria Typical accuracy of the recovery of the drug substance in the mixture is expected to be about 98 to 102%. MOKARAM HOSSAIN 30-Nov-24 21  Procedure  Precision is usually investigated at three levels: repeatability, intermediate precision, and reproducibility.  For simple formulation it is important that precision be determined using authentic homogeneous samples. MOKARAM HOSSAIN 30-Nov-24 22  A justification will be required if a homogeneous sample is not possible and artificially prepared samples or sample solutions are used. MOKARAM HOSSAIN 30-Nov-24 23  Procedure  A minimum of nine determinations covering the specified range for the procedure (e.g., three concentrations/three replicates as in the accuracy experiment), or  A minimum of six determinations at 100% of the test concentration. MOKARAM HOSSAIN 30-Nov-24 24  The standard deviation, relative standard deviation (coefficient of variation), and confidence interval should be reported as required by the ICH. MOKARAM HOSSAIN 30-Nov-24 25  Procedure  Typical parameters that are investigated include day-to-day variation, analyst variation, and equipment variation.  The ICH recommended the reporting of standard deviation, relative standard deviation (coefficient of variation), and confidence interval of the data. MOKARAM HOSSAIN 30-Nov-24 26  Procedure  similar studies need to be performed at other laboratories using the same homogeneous sample lot and the same experimental design. MOKARAM HOSSAIN 30-Nov-24 27  Common variations that are investigated for robustness include filter effect, stability of analytical solutions, extraction time during sample preparation, pH variations in the mobile- phase composition, variations in mobile-phase composition, columns, temperature effect, and flow rate. MOKARAM HOSSAIN 30-Nov-24 28 Results from Method Transfer between Two Laboratories MOKARAM HOSSAIN 30-Nov-24 29 Stability of Sample and Standard Solutions MOKARAM HOSSAIN 30-Nov-24 30 Effect of Filter MOKARAM HOSSAIN 30-Nov-24 31  Procedure  The specificity of a test method is determined by comparing test results from an analysis of samples containing impurities, degradation products, or placebo ingredients with those obtained from an analysis of samples without impurities, degradation products, or placebo ingredients. MOKARAM HOSSAIN 30-Nov-24 32  For the purpose of a stabilityindicating assay method, degradation peaks need to be resolved from the drug substance.  Critical separations in chromatography should be investigated at the appropriate level.  Specificity can best be demonstrated by the resolution of two chromographic peaks that elute close to each other. MOKARAM HOSSAIN 30-Nov-24 33 Overlay chromatogram of an impurity solution with a sample solution. MOKARAM HOSSAIN 30-Nov-24 34  During optimization of the formulation or drug substance synthetic process, significant changes may have to be introduced into the process.  The composition and/or the final anufacturing process of a sample analyzed with the method have been modified after optimization. MOKARAM HOSSAIN 30-Nov-24 35  The method was found to be deficient in some areas, such as precision and system suitability. This is especially important as the analytical laboratory gets more experience and more information as to the degradation profile of the sample as it progresses toward submission. If a new impurity is found that makes the method deficient, this method will need to be revalidated. MOKARAM HOSSAIN 30-Nov-24 36  Changes in equipment or in suppliers of critical supplies at the time of manufacturing. This is important, as critical components of the manufacturing process have the potential to change the degradation profile of the product. MOKARAM HOSSAIN 30-Nov-24 37  HPLC Instrumentation Errors  Qualification of Instruments The status of the qualification of HPLC and other equipment used for the analytical procedure must always be checked. MOKARAM HOSSAIN 30-Nov-24 38  HPLC Instrumentation Errors  Vacuum Filtering of Mobile Phase Vacuum filtering of the mobile phase should be avoided in a procedure that is very ensitive to the level of the organic in the mobile phase. Vacuum suction will evaporate the volatile organic portion during filtration (e.g., acetonitrile or methanol), and may lead to variation of the chromatography. MOKARAM HOSSAIN 30-Nov-24 39  Procedural Errors  Expiry of Mobile Phase  Use of Ion-Pairing Reagents in Mobile Phase  Quantitation of Salts (e.g., Hydrochloride and Sodium Salt)  Stability of Standard and Sample Solutions  Dilution during Sample and Standard Preparation. MOKARAM HOSSAIN 30-Nov-24 40  Procedural Errors  Range in Validation of Linearity Is Smaller Than Precision and Accuracy.  Dilution during Sample and Standard Preparation. MOKARAM HOSSAIN 30-Nov-24 41  Miscellaneous Errors  Validation Protocol  Acceptance Criteria for Validation Parameter  Documentation of Observation  Absorbance of Analyte. MOKARAM HOSSAIN 30-Nov-24 42 THANK YOU MOKARAM HOSSAIN 30-Nov-24 43