Quality Control of Pharmaceuticals PDF

College of Pharmacy-Alamein Campus The Arab Academy for Science and Technology and Maritime Transport (AASTMT CPS 414: Quality Control of Pharmaceuticals Analytical method validation Presented by: Profess...

College of Pharmacy-Alamein Campus The Arab Academy for Science and Technology and Maritime Transport (AASTMT CPS 414: Quality Control of Pharmaceuticals Analytical method validation Presented by: Professor Dr. Hadir Maher Shalaby Professor of Pharmaceutical Analytical Chemistry Faculty of Pharmacy, Alexandria University, Egypt Content What is method validation? Why is validation important? Steps in method validation Parameters of Analytical Method Validation Revalidation of analytical methods 2 Prof. Dr. Hadir Maher  What is Method Validation? Method Validation if the The word validation simply process of ‘establishing means assessment of validity or documented evidence’ which action of proving effectiveness. provides high degree of Validation is a team effort assurance that the method will where it involves people from meet the requirements for the various disciplines of the plant. intended application. 3 Prof. Dr. Hadir Maher Why is validation important? 4 Prof. Dr. Hadir Maher Regulation agencies for analytical method validation 5 Prof. Dr. Hadir Maher Types of analytical procedures to be validated Identification tests Quantitative tests for impurities content Limit tests for the control of impurities Quantitative tests of the active moiety in samples of drug substance (raw material), finished product or other selected. 6 Prof. Dr. Hadir Maher Steps in Method Validation 7 Prof. Dr. Hadir Maher ICH validation guidelines 8 Prof. Dr. Hadir Maher  USP validation guidelines According to USP, different test methods require different validation schemes. Category I Analytical methods for quantitation of major components of bulk drug substances or active ingredients in finished pharmaceutical products. Category II Analytical methods for the determination of impurities in bulk drug substances or degradation compounds in finished pharmaceutical products. Category III Analytical methods for determination of performance characteristics, e.g. dissolution, performance characteristics, drug release, ……. Category IV Identification tests. 9 Prof. Dr. Hadir Maher Analytical parameters needed for method validation as described in the USP General Chapter , ICH guidelines, and FDA Reviewer Guidance are summarized in the following table: 10 Prof. Dr. Hadir Maher 11 Prof. Dr. Hadir Maher ◼ Accuracy and precision Accurate & precise Accurate & imprecise Inaccurate & Inaccurate & precise imprecise Prof. Dr. Hadir Maher 12  1- Accuracy: ICH/USP ◼ Closeness of the test results obtained by the method to the true value. ◼ Trueness Applicability Accuracy is established for a procedure used for: Assay of drug substance, Assay of drug product, Quantitation of impurities or degradation products Prof. Dr. Hadir Maher 13  Expression of accuracy % Recovery=(Observed/true)x100 Prof. Dr. Hadir Maher 14  Procedure guidelines and acceptance ❖ Procedure guidelines criteria ICH Guidelines recommend assessment of accuracy using minimum of 9 determinations over a minimum of 3 concentration levels covering the specified range (e.g., 3 concentrations/3 replicates each of the total analytical procedure). ❖ Acceptance criteria Accuracy should be reported as percent recovery by the assay of known added amount of analyte in the sample or as the difference between the mean and the accepted true value (absolute error or relative error) at THREE concentration levels (i.e., three concentration levels and three replicates at each level of the total analytical procedure). Prof. Dr. Hadir Maher 15  ICH Accuracy: Drug Substance 1. Application of procedure to an analyte of known concentration (reference material). 2. Comparison of results between proposed method and second well characterized procedure (of known accuracy). Prof. Dr. Hadir Maher 16  ICH Accuracy: Drug Product 1. Application of procedure to synthetic mixture of drug products components (placebo) to which known amounts of drug substance have been added….Spiking. 2. Adding known amounts of drug substance to drug product and assaying by method….Standard addition method. 3. Comparison of results between proposed method and second well characterized procedure (of known accuracy). Prof. Dr. Hadir Maher 17  ICH Accuracy: Impurity Assay 1. Application of procedure of drug substance or drug product samples spiked with known amounts of impurities. 2. Comparison of results between proposed method and second well characterized procedure (of known accuracy). Prof. Dr. Hadir Maher 18  2-Precision: ICH/USP Ball Ball Strike Strike Ball Strike ◼ The closeness of agreement (degree of Ball Ball Ball StrikeStrike Ball Ball Strike scatter) between a series of measurements 218330_2 obtained from multiple samplings of the same homogeneous sample. Expressed as: Variance, standard deviation, relative standard deviation Prof. Dr. Hadir Maher 19 Precision parameters: Standard Sample Result Average Variance Deviation %CV =AVERAGE(D5: =100*(StDev/Avera D8) =VAR(D5:D8) =STDEV(D5:D8) ge) 1.1 0.12 0.1475 0.00383 0.061847 41.929888 1.2 0.24 1.3 0.11 1.4 0.12 2.1 0.3 0.31 0.00047 0.021602 6.9685384 2.2 0.34 2.3 0.29 2.4 0.31 3.1 0.52 0.48 0.003 0.054772 11.410887 3.2 0.51 3.3 0.49 3.4 0.4 Prof. Dr. Hadir Maher 20  ICH Precision… Considered at 3 Levels ◼ Repeatability (intra-assay precision) ◼ Intermediate Precision (variability within a laboratory) ◼ Reproducibility (precision between laboratories) Prof. Dr. Hadir Maher 21  ICH Intra-assay a) Repeatability Intra-day ◼ Express the precision under Should be assessed the same operating conditions using minimum of 9 determinations over a short interval of (3 concentrations/ 3 time. replicates) or ◼ Also referred to as Intra- Minimum of 6 assay precision determinations at the 100% level. Prof. Dr. Hadir Maher 22  ICH Within-lab b) Intermediate Precision Express within-laboratory Depends on the circumstances Inter-day variations. under which the procedure is Expressed in terms of intended to be used. standard deviation, relative Studies should include varying days, standard deviation (coefficient analysts, equipment, etc. of variation) and confidence interval. Prof. Dr. Hadir Maher 23  ICH c) Reproducibility Reproducibility expresses the precision between laboratories. The ICH recommends that reproducibility studies for inclusion of procedures to PHARMACOPOEIAS ◼ Collaborative studies, usually applied to standardisation of methodology ◼ Transfer of technology ◼ Compendial methods Prof. Dr. Hadir Maher 24 Random (precision) and systematic (accuracy) deviations precise not precise and but correct correct precise not precise but and wrong wrong Prof. Dr. Hadir Maher 25  3-Linearity: ICH/USP ◼ Ability of an assay to elicit a direct and proportional response to changes in analyte concentration. Prof. Dr. Hadir Maher 26  Linearity Should be Evaluated 146029_2 ◼ By Visual Inspection of plot of signals vs. analyte concentration Requires a minimum of 5 concentration levels ◼ By Appropriate statistical methods ◼ Linear Regression (y = mx + b) ◼ Correlation Coefficient, y-intercept (b), slope (m), residual sum of squares ❖ Acceptance Criteria The correlation coefficient will be >0.99.. For impurities, the correlation coefficient will be > 0.98, Prof. Dr. Hadir Maher 27  4-Range: USP/ICH ◼ The interval between the upper and lower concentrations of analyte in the sample that have been demonstrated to have a suitable level of precision, accuracy, and linearity. Prof. Dr. Hadir Maher 28  4-Range: USP/ICH ◼ Normally derived from Linearity studies. ◼ Established by confirming that the method provides acceptable degree of linearity, accuracy, and precision. ◼ Specific range dependent upon intended application of the procedure. Prof. Dr. Hadir Maher 29  Minimum Specified Range: ◼ For Drug Substance & Drug product Assay ◼ 80 to 120% of test Concentration ◼ For Content Uniformity Assay ◼ 70 to 130% of test Concentration ◼ For Dissolution Test Method ◼ +/- 20% over entire Specification Range ◼ For Impurity Assays ◼ From Reporting Level to 120% of Impurity Specification for Impurity Assays ◼ From Reporting Level to 120% of Assay Specification for Impurity/Assay Methods Prof. Dr. Hadir Maher 30  5-Specificity ◼ The terms specificity and selectivity are often used Inter- changeably. ◼ SPECIFICITY: ICH defines specificity as the ability to measure accurately the analyte in the presence of other components (other active ingredients, excipients, impurities, and degradation products) ◼ SELECTIVITY: of a method is a measure of the extent to which the method can determine a particular compound in the analyzed matrices without interference from matrix components. Prof. Dr. Hadir Maher 31  Specificity: ICH/USP ◼ An investigation of specificity should be conducted during the validation of an identification test, an impurities assay, and a potency assay. ◼ Procedures used will depend on the intended objective of the analytical procedure. Prof. Dr. Hadir Maher 32  ICH Specificity: Identification ◼ Should be able to discriminate between compounds closely related in structure. ◼ Confirmed by obtaining negative results for samples with spiked related compounds and positive results for samples with analyte. ◼ Choice of potential interfering substances should be based on sensible scientific judgment considering substances that could likely occur. Prof. Dr. Hadir Maher 33 Specificity: Potential Interference ◼ Placebo ◼ Impurities ◼ Other active ingredients ◼ Drug Substance Degradants ◼ Drug Product Degradants ◼ Related Substances ◼ Packaging Extractables Prof. Dr. Hadir Maher 34  ICH Specificity: Impurities/Assay Chromatographic Methods Demonstrate Resolution ◼ Impurities/Degradants Available ◼ Spike with impurities/degradants ◼ Show resolution and a lack of interference ◼ Impurities/Degradants Not Available ◼ Stress Samples ◼ For assay, Stressed and Unstressed Samples should be compared. ◼ For impurity test, impurity profiles should be compared. ❖ Acceptance limit: Base line resolution from any peak in the chromatogram is > 1.5. Prof. Dr. Hadir Maher 35 Peak purity tests can also be evaluated with: The spectra of Photodiode array detectors Mass spectrometry Prof. Dr. Hadir Maher 36  Specificity, practical evaluation Forced Degradation Studies Heat 216075_2 216076_2 ◼ ◼ Humidity ◼ Acid Hydrolysis ◼ Base Hydrolysis 1966_2 ◼ Oxidation Peak purity tests can also be evaluated with: 146033_2 ◼ Light The spectra of Photodiode array detectors Mass spectrometry Prof. Dr. Hadir Maher 37  Representative HPLC chromatograms of impurity-spiked solution, acid degradation, base degradation and oxidative degradation Prof. Dr. Hadir Maher 38 Prof. Dr. Hadir Maher 39  Specificity, practical evaluation ◼ Specificity: Overlay chromatogram of an impurity solution with a sample solution From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons Prof. Dr. Hadir Maher 40  6-Detection limit (means) Is any of it present? Is it there? ◼ Lowest amount of analyte in a sample that can be detected but not necessarily Prof. Dr. Hadir Maher quantitated. 41  7-Quantitation limit How much of it is present??? How much of it is there? ◼ Lowest amount of analyte in a sample that can be quantified with suitable accuracy and precision. The quantitation limit is used particularly for the determination of impurities and/or degradation products Prof. Dr. Hadir Maher 42  ICH Detection Limit (DL, LOD) and Quantitation Limit (QL, LOQ) Estimated by 1. Based in Visual Evaluations - Used for non-instrumental methods 2. Based on Signal-to Noise-Ratio - 3:1 for Detection Limit - 10:1 for Quantitation Limit 3. Based on Standard Deviation of the Response and the Slope Prof. Dr. Hadir Maher 43  ❖Procedures The ICH approaches are: 1. Based on Visual Evaluation: Practically, LOD, can be measured by the serial dilution of samples until the peak can no longer be observed. For LOQ can be determined by analyzing successively diluted samples until the requisite levels of accuracy and precision are achieved. Prof. Dr. Hadir Maher 44 2- Based on Signal-to Noise-Ratio LOD, LOQ and Signal to Noise Ratio (SNR) LOQ Signal to Noise = 10:1 LOD Signal to Noise = 3:1 Noise Prof. Dr. Hadir Maher 45 3- Based on standard deviation of the response and slope 3.3s 10s DL = QL = S S ◼ S = slope of calibration curve ◼ s = standard deviation of blank readings or standard deviation of regression line Prof. Dr. Hadir Maher 46  ICH 8- Robustness ◼ Definition: Capacity to remain unaffected by small but deliberate variations in method parameters and provide an indication of its reliability during normal usage. ◼ Determination: Comparison results under differing conditions with precision under normal conditions ◼ Variations may include: stability of analytical solution, variation of pH in a mobile phase, different column (lot/supplier), temperature, flow rate. Prof. Dr. Hadir Maher 47 Robustness Variations Prof. Dr. Hadir Maher 48  9-ICH/USP System Suitability ◼ ICH ◼ Definition: evaluation of equipment, electronic, analytical operations and samples as a whole ◼ Determination: repeatability, tailing factor (T), capacity factor (k’), resolution (R), and theoretical Plates (N) Prof. Dr. Hadir Maher 49 ◼ USP 23 ◼ System Suitability Requirements Parameters Recommendations K’ In general k’ ≥ 2.0 R > 2, between the peak of interest and the closest potential interferent R (degradant, internal STD, impurity, excipient, etc…..) T T≤2 N In general N > 2000 Repeatability RSD ≤ 2.0% (n ≥ 5) Prof. Dr. Hadir Maher 50  RUGGEDNESS* ❖ Definition It is a measure of reproducibility of test results under the variation in conditions normally expected from laboratory to laboratory and from analyst to analyst. It is the degree of reproducibility of test results obtained by the analysis of the same samples under a variety of conditions, such as: ✓ Different laboratories. ✓ Different analysts. ✓ Different instruments. ✓ Different lots of reagents. ✓ Different days, etc. ❖ Procedures It is determined by analysis of aliquots from homogeneous lots in different laboratories, by different analysts, using operational and environmental conditions that may differ but still within the specified parameters of the assay. *This item is not included in ICH guidelines Prof. Dr. Hadir Maher 51 Robustness Prof. Dr. Hadir Maher 52  METHOD REVALIDATION As the method under validation is required to be used as written, any modification requires revalidation. Validation should be in the form of revalidation with the following method changes. For Chromatographic Methods, significant changes include: 1) Changes in the product for which the method was validated. 2) Use of the assay for a product different from that for which it was validated. 3) Instrumental changes. 4) Reagent changes. 5) Procedural changes. 6) Technological changes (e.g., developments in column and/ or instrumental technology). Prof. Dr. Hadir Maher 53  More specific recommendations for which method parameters should be revalidated: Important Prof. Dr. Hadir Maher 54 Prof. Dr. Hadir Maher 55