Validation Part4
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The document discusses analytical method validation for good manufacturing practices. It covers validation of pharmacopoeial and non-pharmacopoeial methods, and characteristics that should be considered during validation including accuracy, precision, robustness, specificity, linearity, range, detection limit and quantitation limit. The objectives are to validate analytical methods to demonstrate suitability for their intended purpose and ensure results are reliable, accurate and reproducible.
Validation Part4
- 1. Supplementary Training Modules on Good Manufacturing Practice Validation WHO Technical Report Series, No. 937, 2006. Annex 4. Validation | Slide 1 of 27 August 2006
- 2. Validation Part 1. General overview on qualification and validation Part 2. Qualification of HVAC and water systems Part 3. Cleaning validation Part 4. Analytical method validation Part 5. Computerized system validation Part 6. Qualification of systems and equipment Part 7. Non sterile product process validation Validation | Slide 2 of 27 August 2006
- 3. Supplementary Training Modules on Good Manufacturing Practice Analytical Method Validation Part 4 WHO Technical Report Series, No. 937, 2006. Annex 4 Appendix 4 Validation | Slide 3 of 27 August 2006
- 4. Validation Objectives To discuss various aspects of analytical method validation including: Principles of analytical method validation Pharmacopoeia methods Non-pharmacopoeia methods Approaches to analytical method validation Characteristics of analytical procedures Validation | Slide 4 of 27 August 2006
- 5. Validation Principle Guideline presents information on the characteristics to be considered (other approaches may be followed) Manufacturers to demonstrate - analytical procedure is suitable for its intended purpose (validation) Validate analytical methods - whether they indicate stability or not Validated by R&D before being transferred to the quality control unit when appropriate 1.1 – 1.4 Validation | Slide 5 of 27 August 2006
- 6. Validation General Specifications for materials and products, with standard test methods Manufacturer to use “pharmacopoeial specifications and methods”, or suitably developed “non-pharmacopoeial specifications and methods” as approved by the national drug regulatory authority Use well-characterized reference materials, with documented purity, in the validation study 2.1 – 2.3 Validation | Slide 6 of 27 August 2006
- 7. Validation General (2) Tests include: – identification tests – assay of drug substances and pharmaceutical products – content of impurities and limit tests for impurities – dissolution testing and determination of particle size Results should be reliable, accurate and reproducible 2.4 – 2.5 Validation | Slide 7 of 27 August 2006
- 8. Validation General (3) When should verification or revalidation be done? – changes in the process for synthesis of the drug substance – changes in the composition of the finished product – changes in the analytical procedure – transfer of methods from one laboratory to another – changes in major pieces of equipment instruments Extent depends on the nature of the change(s) Evidence of “analyst proficiency” 2.6 – 2.8 Validation | Slide 8 of 27 August 2006
- 9. Validation Pharmacopoeial/Non-pharmacopoeial methods Pharmacopoeial methods: – prove that the methods are suitable for routine use in the laboratory (verification) – for determination of content or impurities in products, demonstrate that method is specific for the substance under consideration (no placebo interference) Non-pharmacopoeial methods: – Should be appropriately validated 3. – 4. Validation | Slide 9 of 27 August 2006
- 10. Validation Method validation Protocol: includes procedures and acceptance criteria Report: documented results Justification needed when non-pharmacopoeial methods are used (if pharmacopoeial methods are available). Justification to include data, e.g. comparisons with the pharmacopoeial or other methods Detailed standard test methods include: – chromatographic conditions, reagents and others 5.1 – 5.3 Validation | Slide 10 of 27 August 2006
- 11. Validation Characteristics that should be considered during validation of analytical methods include: – specificity – linearity – range – accuracy – precision – detection limit – quantitation limit – robustness 6.1 Validation | Slide 11 of 27 August 2006
- 12. Validation Accuracy: is the degree of agreement of test results with the true value, or the closeness of the results obtained by the procedure to the true value. It is normally established on samples of the material to be examined that have been prepared to quantitative accuracy. Accuracy should be established across the specified range of the analytical procedure. Note: it is acceptable to use a “spiked” Accurate but placebo where a known quantity or concentration of a reference material is imprecise used. 6.1.1 Validation | Slide 12 of 27 August 2006
- 13. Validation Precision: is the degree of 6.1.2 agreement among individual results. The complete procedure should be applied repeatedly to separate, identical samples drawn from the same homogeneous batch of material. It should be measured by the scatter of individual results from the mean (good grouping) and expressed as the relative standard deviation (RSD). Inaccurate but precise Validation | Slide 13 of 27 August 2006
- 14. Validation Precision: Repeatability: A minimum of nine determinations covering the specified range for the procedure, e.g. – three concentrations/three replicates each, or a minimum of six determinations at 100% of the test concentration Intermediate precision: Within-laboratory variations – usually on different days, different analysts and different equipment. (If reproducibility is assessed, a measure of intermediate precision is not required.) Reproducibility: Precision between laboratories 6.1.2.1 – 6.1.2.3 Validation | Slide 14 of 27 August 2006
- 15. Validation Accurate and Precise Validation | Slide 15 of 27 August 2006
- 16. Validation Robustness (or ruggedness): is the ability of the procedure to provide analytical results of acceptable accuracy and precision under a variety of conditions. The results from separate samples are influenced by changes in the operational or environmental conditions. Robustness should be considered during the development phase, and should show the reliability of an analysis when deliberate variations are made in method parameters. 6.1.3 Validation | Slide 16 of 27 August 2006
- 17. Validation Factors that can have an effect on robustness when performing chromatographic analysis include: – stability of test and standard samples and solutions – reagents (e.g. different suppliers) – different columns (e.g. different lots and/or suppliers) – extraction time – variations of pH of a mobile phase – variations in mobile phase composition – temperature – flow rate 6.1.3.1 Validation | Slide 17 of 27 August 2006
- 18. Validation Linearity: indicates the ability to produce results that are directly proportional to the concentration of the analyte in samples – A series of samples should be prepared in which the analyte concentrations span the claimed range of the procedure. If there is a linear relationship, test results should be evaluated by appropriate statistical methods – A minimum of five concentrations should be used 6.1.4 Validation | Slide 18 of 27 August 2006
- 19. Validation Calculated analyte in mg/mL 0.040 Linearity 0.035 of an analyte in a material Table of values (x,y) 0.030 x y 0.025 Reference Calculated # material mg/ml mg/ml 0.020 1 0.0100 0.0101 0.015 Reference material mg/ml 2 0.0150 0.0145 0.010 3 0.0200 0.0210 0.01 0.015 0.02 0.025 0.03 0.035 0.04 4 0.0250 0.0260 5 0.0300 0.0294 6 0.0400 0.0410 Validation | Slide 19 of 27 August 2006
- 20. Validation Range: is an expression of the lowest and highest levels of analyte that have been demonstrated to be determinable for the product The specified range is normally derived from linearity studies 6.1.5 Validation | Slide 20 of 27 August 2006
- 21. Validation Specificity (selectivity): is the ability to measure unequivocally the desired analyte in the presence of components such as excipients and impurities that may also be expected to be present. An investigation of specificity should be conducted during the validation of identification tests, the determination of impurities and assay 6.1.6 Validation | Slide 21 of 27 August 2006
- 22. Validation Detection limit (limit of detection): is the smallest quantity of an analyte that can be detected, and not necessarily determined, in a quantitative fashion Approaches (instrumental or non-instrumental): – visual evaluation – signal to noise ratio – standard deviation of the response and the slope – standard deviation of the blank – calibration curve 6.1.7 Validation | Slide 22 of 27 August 2006
- 23. Validation Quantitation limit (limit of quantitation): is the lowest concentration of an analyte in a sample that may be determined with acceptable accuracy and precision Approaches (instrumental or non-instrumental): – visual evaluation – signal to noise ratio – standard deviation of the response and the slope – standard deviation of the blank – calibration curve 6.1.8 Validation | Slide 23 of 27 August 2006
- 24. Validation LOQ, LOD and SNR Limit of Quantitation Limit of Detection Peak B LOQ Signal to Noise Ratio Peak A LOD Baseline noise Validation | Slide 24 of 27 August 2006
- 25. Validation Table 1. Characteristics to consider during analytical validation 6.2 Validation | Slide 25 of 27 August 2006
- 26. Validation System suitability testing Integral part of many analytical procedures Based on the concept that: – the equipment, electronics, analytical operations and samples to be analysed constitute an integral system that can be evaluated as such Depends on the type of procedure being evaluated – e.g. resolution test for an HPLC procedure 6.3 Validation | Slide 26 of 27 August 2006
- 27. Validation Group session Validation | Slide 27 of 27 August 2006
Editor's Notes
- #2: दिसंबर 4, 2012 In this supplementary training module, we will be looking at the recommendations by WHO, on Validation and qualification. The module consists of 7 parts: Part 1. General overview on qualification and validation Part 2. Qualification of HVAC and water systems Part 3. Cleaning validation Part 4. Analytical method validation Part 5. Computerized system validation Part 6. Qualification of systems and equipment Part 7. Non sterile product process validation Each part deals with a specific topic, and each part can be presented in about one to one and a half hours time. Presenters should know the topics and add practical examples to the texts taken from the WHO guideline.
- #3: दिसंबर 4, 2012
- #4: दिसंबर 4, 2012
- #5: दिसंबर 4, 2012
- #6: दिसंबर 4, 2012 1. Principle 1.1 This appendix presents some information on the characteristics that should be considered during validation of analytical methods. Approaches other than those specifi ed in this appendix may be followed and may be acceptable. Manufacturers should choose the validation protocol and procedures most suitable for testing of their product. 1.2 The manufacturer should demonstrate (through validation) that the analytical procedure is suitable for its intended purpose. 1.3 Analytical methods, whether or not they indicate stability, should be validated. 1.4 The analytical method should be validated by research and development before being transferred to the quality control unit when appropriate.
- #7: दिसंबर 4, 2012 2. General 2.1 There should be specifi cations for both, materials and products. The tests to be performed should be described in the documentation on standard test methods. 2.2 Specifi cations and standard test methods in pharmacopoeias (“pharmacopoeial methods”), or suitably developed specifi cations or test methods (“non-pharmacopoeial methods”) as approved by the national drug regulatory authority may be used. 2.3 Well-characterized reference materials, with documented purity, should be used in the validation study.
- #8: दिसंबर 4, 2012 2.4 The most common analytical procedures include identifi cation tests, assay of drug substances and pharmaceutical products, quantitative tests for content of impurities and limit tests for impurities. Other analytical procedures include dissolution testing and determination of particle size. 2.5 The results of analytical procedures should be reliable, accurate and reproducible. The characteristics that should be considered during validation of analytical methods are discussed in paragraph 6.
- #9: दिसंबर 4, 2012 2.6 Verifi cation or revalidation should be performed when relevant, for example, when there are changes in the process for synthesis of the drug substance; changes in the composition of the fi nished product; changes in the analytical procedure; when analytical methods are transferred from one laboratory to another; or when major pieces of equipment instruments change. 2.7 The verifi cation or degree of revalidation depend on the nature of the change(s). 2.8 There should be evidence that the analysts, who are responsible for certain tests, are appropriately qualifi ed to perform those analyses (“analyst profi ciency”).
- #10: दिसंबर 4, 2012 3. Pharmacopoeial methods 3.1 When pharmacopoeial methods are used, evidence should be available to prove that such methods are suitable for routine use in the laboratory (verifi cation). 3.2 Pharmacopoeial methods used for determination of content or impurities in pharmaceutical products should also have been demonstrated to be specifi c with respect to the substance under consideration (no placebo interference). 4. Non-pharmacopoeial methods 4.1 Non-pharmacopoeial methods should be appropriately validated.
- #11: दिसंबर 4, 2012 5. Method validation 5.1 Validation should be performed in accordance with the validation protocol. The protocol should include procedures and acceptance criteria for all characteristics. The results should be documented in the validation report. 5.2 Justifi cation should be provided when non-pharmacopoeial methods are used if pharmacopoeial methods are available. Justifi cation should include data such as comparisons with the pharmacopoeial or other methods. 5.3 Standard test methods should be described in detail and should provide suffi cient information to allow properly trained analysts to perform the analysis in a reliable manner. As a minimum, the description should include the chromatographic conditions (in the case of chromatographic tests), reagents needed, reference standards, the formulae for the calculation of results and system suitability tests.
- #12: दिसंबर 4, 2012
- #13: दिसंबर 4, 2012 6.1.1 Accuracy is the degree of agreement of test results with the true value, or the closeness of the results obtained by the procedure to the true value. It is normally established on samples of the material to be examined that have been prepared to quantitative accuracy. Accuracy should be established across the specifi ed range of the analytical procedure. Note: it is acceptable to use a “spiked” placebo where a known quantity or concentration of a reference material is used.
- #14: दिसंबर 4, 2012 6.1.2 Precision is the degree of agreement among individual results. The complete procedure should be applied repeatedly to separate, identical samples drawn from the same homogeneous batch of material. It should be measured by the scatter of individual results from the mean (good grouping) and expressed as the relative standard deviation (RSD).
- #15: दिसंबर 4, 2012 6.1.2.1 Repeatability should be assessed using a minimum of nine determinations covering the specifi ed range for the procedure e.g. three concentrations/ three replicates each, or a minimum of six determinations at 100% of the test concentration. 6.1.2.2 Intermediate precision expresses within-laboratory variations (usually on different days, different analysts and different equipment). If reproducibility is assessed, a measure of intermediate precision is not required. 6.1.2.3 Reproducibility expresses precision between laboratories.
- #16: दिसंबर 4, 2012
- #17: दिसंबर 4, 2012 6.1.3 Robustness (or ruggedness ) is the ability of the procedure to provide analytical results of acceptable accuracy and precision under a variety of conditions. The results from separate samples are infl uenced by changes in the operational or environmental conditions. Robustness should be considered during the development phase, and should show the reliability of an analysis when deliberate variations are made in method parameters.
- #18: दिसंबर 4, 2012 6.1.3.1 Factors that can have an effect on robustness when performing chromatographic analysis include: — stability of test and standard samples and solutions; — reagents (e.g. different suppliers); — different columns (e.g. different lots and/or suppliers); — extraction time; — variations of pH of a mobile phase; — variations in mobile phase composition; — temperature; and — flow rate.
- #19: दिसंबर 4, 2012 6.1.4 Linearity indicates the ability to produce results that are directly proportional to the concentration of the analyte in samples. A series of samples should be prepared in which the analyte concentrations span the claimed range of the procedure. If there is a linear relationship, test results should be evaluated by appropriate statistical methods. A minimum of fi ve concentrations should be used.
- #20: दिसंबर 4, 2012 This is a typical plot of raw data to check for linearity and range. The table of raw data on the right hand side has been plotted using linear regression to obtain a line of best fit. The x axis is the reference material that was weighed out and the y axis the calculated values from the instrument readings. These are plotted (the blue line with the points as a yellow box and error bars) and a linear regression analysis is performed. The two yellow lines are the 2-sided 95% confidence limits.
- #21: दिसंबर 4, 2012 Range : is an expression of the lowest and highest levels of analyte that have been demonstrated to be determinable for the product . The specified range is normally derived from linearity studies.
- #22: दिसंबर 4, 2012 6.1.6 Specificity ( selectivity ) is the ability to measure unequivocally the desired analyte in the presence of components such as excipients and impurities that may also be expected to be present. An investigation of specifi city should be conducted during the validation of identification tests, the determination of impurities and assay.
- #23: दिसंबर 4, 2012 6.1.7 Detection limit ( limit of detection ) is the smallest quantity of an analyte that can be detected, and not necessarily determined, in a quantitative fashion. Approaches may include instrumental or non-instrumental procedures and could include those based on: — visual evaluation; — signal to noise ratio; — standard deviation of the response and the slope; — standard deviation of the blank; and — calibration curve.
- #24: दिसंबर 4, 2012 6.1.8 Quantitation limit ( limit of quantitation ) is the lowest concentration of an analyte in a sample that may be determined with acceptable accuracy and precision. Approaches may include instrumental or non-instrumental procedures and could include those based on: — visual evaluation; — signal to noise ratio; — standard deviation of the response and the slope; 140 — standard deviation of the blank; and — calibration curve.
- #25: दिसंबर 4, 2012 There are no specific criteria set for the Limit of Quantitation (LOQ) and Limit of Detection (LOD) but guidance is available from specifications and pharmacopeias . The noise is measured by running the instrument at maximum gain with no test being processed. The ripple generated is noise due to the instrument’s electronics, etc. The peak is measured relative to this noise and the ratio is calculated. This is known as the Signal to Noise Ratio (SNR ). Generally : LOD SNR should be greater than 2:1. Peak A is acceptable for LOD but not for quantitation ; The LOD can be calculated if the standard deviation (SD) of the response (which is standard deviation of the blank) and the slope is determined : LOD = 3.3 x SD slope Similarly LOQ = 10 x SD slope The LOQ SNR should generally be above 10:1. Peak B is suitable for quantitation ; Precision as a percentage relative standard deviation should be 5 - 10% at the limit for LOQ .
- #26: दिसंबर 4, 2012
- #27: दिसंबर 4, 2012 6.3 System suitability testing System suitability testing is an integral part of many analytical procedures. The tests are based on the concept that the equipment, electronics, analytical operations and samples to be analysed constitute an integral system that can be evaluated as such. System suitability test parameters that need to be established for a particular procedure depend on the type of procedure being evaluated, for instance, a resolution test for an HPLC procedure.
- #28: दिसंबर 4, 2012 The trainer should give the participants a case study, based on experience