Analytical method validation

describes the parameters of validation

1. PREPARED BY: DEVIPRIYA PV M PHARM Department of Pharmaceutical Analysis ANALYTICAL METHOD VALIDATION

2. DEFINITION 2 “ Documented programme which provides a high degree of assurance that an analytical assay method will consistently determine the presence, absence or quality of one or more attributes with accuracy and precision”

3. OBJECTIVE 3 To obtain consistent , reliable and accurate data. The results from method validation can be used to judge the quality, reliability and consistency of analytical results, which is an integral part of any good analytical practice.

4. 4 Analytical methods need to be validated, verified, or revalidated in the following instances: Before initial use in routine testing. When transferred to another laboratory. Whenever the conditions or method parameters for which the method has been validated change and the change is outside the original scope of the method.

5. ICH 5 Requirements: Analytical methods should be validated unless the method employed is included in the relevant pharmacopoeia or other recognized standard reference. The degree of analytical validation performed should reflect the purpose of the analysis and the stage of the API production process. Appropriate qualification of analytical equipment should be considered before starting validation of analytical methods.

6. USP 6

7. PARAMETERS FOR METHOD VALIDATION 7

8. SPECIFICITY 8 Ability to assess unequivocally the analyte in the presence of components which may be expected to be present. Include impurities, degradants , matrix etc. Procedures used to demonstrate specificity: Identification test Assay(content or potency). Purity test.

9. ACCURACY 9 ICH defines the accuracy of an analytical procedure as the closeness of agreement between the conventional true value or an accepted reference value and the value found. The extent to which test results generated by the method and the true value agree. Should be assessed using a minimum of 3 concentration levels, each in triplicate.

10. 10 Should be reported as: Percent recovery of known amount added The difference between the mean assay result and the accepted value.

11. PRECISION 11 The closeness of agreement between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions. Precision may be considered at three levels: Repeatability Intermediate precision Reproducibility

12. LINEARITY 12 ICH defines linearity of an analytical procedure as its ability to obtain test results that are directly proportional to the concentration of analyte in the sample. Evaluated by: Visual inspection of plot. Statistical methods. Acceptance criteria

13. RANGE 13 ICH defines the range of an analytical procedure as the interval from the upper to the lower concentration of analyte in the sample for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity.

14. LIMIT OF DETECTION(LOD) 14 ICH defines the detection limit of an individual analytical procedure as the lowest amount of analyte in a sample which can be detected but not necessarily quantitated as an exact value. Evaluated by: Signal to noise ratio Visual evaluation Standard deviation of the response and the slope.

15. LIMIT OF QUANTITATION(LOQ) 15 ICH defines the LOQ of an individual analytical procedure as the lowest amount of analyte in a sample which can be quantitatively determined with suitable precision and accuracy.

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17. RUGGEDNESS 17 The degree of reproducibility of results obtained under a variety of conditions, such as different laboratories, analysts, instruments, environmental conditions, operators and materials. Expressed as %RSD. Determined by the analysis of aliquots from homogeneous lots in different laboratories.

18. ROBUSTNESS 18 ICH defines the robustness of an analytical procedure as a measure of its capacity to remain unaffected by small, but deliberate variations in method parameters. It provides an indication of the procedure’s reliability during normal usage.

19. METHOD VALIDATION PROCESS 19 The validity of a specific method should be demonstrated in laboratory experiments using samples or standards that are similar to unknown samples analyzed routinely. The process is broken down in phases because of the length of time and complexity.

20. VALIDATION PHASES 20

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22. VALIDATION PLANNING 22 The master plan should include: Purpose and scope. Glossary. Responsibilities, like user departments, management, QA Method performance characteristics and approaches for testing. Steps for method validation. Selection of tests and acceptance criteria. Approach and parameters for system suitability testing.

23. 23 Modification and revalidation methods. Verification of Compendial and standard methods. Transfer of analytical methods. List of required SOPs. Approval process, documentation and archiving. Templates for project plans, test protocols and validation reports.

24. TESTING FOR PERFORMANCE CHARACTERISTICS 24 PREPARATION: Any chemicals used to determine critical validation parameters, should be available in sufficient quantities, accurately identified, sufficiently stable and checked for exact composition and purity according to specifications. Any other materials and consumables, should be new and qualified.

25. 25 Analytical equipment should be clearly defined, well characterized, qualified or calibrated Operators should be sufficiently familiar with the technique and equipment. TEST EXECUTION: Specificity/selectivity Repeatability of retention times and peak areas Linearity, LOQ, LOD , range Accuracy at different concentrations Intermediate precision Reproducibility

26. SELECTING VALIDATION PARAMETERS AND LIMITS 26

27. 27 Validation report and documentation. Method Adjustments. Changes. Revalidation. Verification of Standard and Compendial Methods

28. VALIDATION OF BIOLOGICALS 28 A substance that is produced by or extracted from a biological source and that needs for its characterization and the determination of its quality a combination of physical-chemical-biological testing, together with the production process and its control.

29. CONTENT OF ANALYTICAL PROCEDURES 29 Principle/scope. Apparatus/equipment. Operating parameters. Reagents/standards. Sample preparation. Standards control solution preparation. Procedure. System Suitability. Calculations. Data reporting.

30. 30 For biologicals follow a two-tired approach for reference standard and materials. Comparison of each new reference standard with a primary reference standard so that it is linked to clinical trial material and the current manufacturing process. Statistical analysis of validation data can be used to evaluate validation characteristics against predetermined acceptance criteria.

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