Learn Analytical Method Validation (AMV) step by step with clear guidelines, key parameters, classifications, and expert strategies. A must-read for pharma QA/QC professionals
Analytical method validation (AMV) is the process of generating documented evidence that provides a high level of assurance that a specific process will consistently produce results meeting predefined specifications and quality attributes. This validation encompasses both the analytical method and the equipment used, ensuring reliability, accuracy, and consistency in testing.
AMV is essential in determining the reliability and suitability of an analytical method for its intended purpose. In the pharmaceutical industry, where the quality of drug substances and drug products is critical, validated analytical methods are the cornerstone of ensuring safety, efficacy, and regulatory compliance.
Because analytical methods directly influence product quality decisions, AMV plays a pivotal role across all stages of drug development and manufacturing. However, it is a complex, resource-intensive process that requires significant expertise, careful planning, and in-depth knowledge. Recognising this, I’ve created this guide to simplify the process and share skill-based insights based on practical experience.
In this article, I will walk you through analytical method validation step by step, in a clear and easy-to-understand format.I will cover:
Whether you’re a student, a beginner in QA/QC, or a professional seeking a refresher, this guide is designed to help you grasp the essentials of analytical method validation with confidence.
Analytical method validation is the process of generating documented evidence that provides a high level of assurance that a specific process will consistently produce results meeting predefined specifications and quality attributes. This validation encompasses both the analytical method and the equipment used, ensuring reliability, accuracy, and consistency in testing.
Drug substances that need validation may be final Active Pharmaceutical Ingredients (APIs), Intermediates, Raw materials and Final products. *Generally validation is performed for the following types of methods:
*Note: Based on requirement other methods can be considered for validation
Cleaning validation is the validation of the Analytical method used for cleaning the equipment.
Related Topic: How to perform cleaning validation?
The following guidelines are widely considered in the Pharmaceutical industries for Analytical method validation:
The following 7- Step Strategy should be followed for any Analytical method validation:
The following are the 6-classification of the chromatographic method:
The following Analytical Validation parameters are performed during method validation:
The ability of an analytical method to measure accurately and specifically the analyte in the presence of components that may be expected to be present in the sample matrix, e.g. impurities, degradation products. Specificity is done by spiking the main substance with all identified available impurities. Impurities which are not mentioned in the monograph should also be considered. PDA detectors and mass spectrometers are used to check peak purity. This test is done in all validation methods.
Where a SIM method is validated, the tested material is exposed to stress degradation and stress conditions must include:
The assay, peak purity and impurity profile of each treated sample are tested.
The precision of an analytical procedure expresses the closeness of agreement between a series of
measurements obtained from several samples of the same homogeneous sample under prescribed conditions. Precision is performed for all types of validations. It is subdivided into:
System repeatability is measured by injecting six replicates of standard solutions at nominal concentration.
Acceptance criteria
The analyte is analysed six times at nominal concentration (six preparation and six injections).
In the method, reproducibility samples are analysed by each analytical and QC analyst using different instruments (make) and different columns (lot).
Ideally, three samples are taken and analysis is performed in duplicate. If three samples are not available, then two sample are taken and one sample is analysed four times, and the other sample is analysed in duplicate. If only one sample is available the analysis is performed six times.
In Intermediate precision, samples are analysed by two different analysts of same lab using different instruments (make) and different columns (lot).
Acceptance criteria: Same as Reproducibility test
Related: How to Perform Precision in Analytical Method Validation?
Accuracy measures the exactness of the analytical method. It is one of the most critical parameters in
Analytical method validation. It is determined by:
Note: Deviation from linearity and Recovery are performed for drug substances/their stages but for dosage forms, only recovery is performed.
Related Topic: How To Perform Accuracy In Analytical Method Validation: Calculation and Case Studies
The lowest concentration of analyte in a sample that can be detected, but not necessarily quantitated as an exact value. It is determined in all types of impurity profile methods including TLC.
The following approaches may be used to calculate DL:
S may be determined from the calibration curve of the analyte.
The σ may be calculated in the following ways:
The lowest concentration of analyte in a sample that can be determined with acceptable precision and accuracy under stated experimental conditions.
The following approaches may be used to calculate DL:
Linearity is a test that shows the ability to obtain test results that are directly, or by a well-defined
mathematical transformation, proportional to the concentration of analyte in samples within a given range. It is performed on all methods in which standards are used and area per cent methods for final material. A minimum of five standard solutions from two stock solutions are injected once. Standard solutions of at least 50 to 150% of nominal concentration are used for Category I (Assay) analysis.
Acceptance Criteria:
R2 should be less than 0.997 for cat. I analysis and R2 should not be less than 0.990 for Cat. II analysis.
The relative response factor shows the ratio of absorbance between two analytes. It is determined by dividing the slope of specified available impurities by the slope of the reference
substance. Each linearity curve must follow linearity acceptance criteria. It is determined for all available specified ou unspecified impurities of final material.
Related Topic:
The measure of the capability of the method to remain unaffected by small deliberate variations in the method parameters during normal usage.
Typical variations are:
Related Topic: How to Perform Robustness in Analytical Method Validation?
The standard/sample solution is tested for up to 5 days by comparison of the response factor of an old solution (injected as a sample) to freshly prepared standard solutions.
The test for standard solution stability is also valid for sample solution stability because same diluent is used for sample-solution preparations
Acceptance criteria: The test passess the acceptance criteria if % RF difference between freshly prepared standard solution and stability solution is ≤ 2%.
The sample solution is tested for up to 5 days for related substances stability testing. Result of old samples solution (stability solution) is compared with the freshly prepared sample solutions.
Acceptance criteria: No new peak higher or equal to QL should appear in the old sample solutions
Related Topic: How To Perform Solution Stability In Analytical Method validation?
Analytical Method Validation protocol is the manual of any Analytical validation work and it is Analytical method specific . It is a pre-approved document that outlines the process for performing analytical method validation. It also includes the purpose, scope and responsibilities of various individuals/departments involved in carrying out the validation.The sensitivity of the method should be considered while designing the protocol to
avoid any surprises or re-work. It is written by the project coordinator and approved by the Analytical Research Manager, Quality Control Manager and Quality Assurance Manager (There may be some changes in the approval process as per the organisation’s policy).
The following are the main sections of Validation protocol:
Once validation work is completed Analytical validation report is prepared based on actual validation work and gets approved by concerned personnel. The following are the important sections of any validation report:
All limitations observed in robustness testing (such as pH, solvent quality, temperature, buffer concentration etc) during validation should be recorded in the validation report and accordingly monograph should also be updated. If an equivalent column is used in the validation, it will also be updated in the standard test procedure and in the monograph
Related Topic: What Is DMF (Drug Master File): Learn In 11 Minutes
Different regulatory agencies provide their own guidelines for analytical method validation, and individual pharmaceutical companies often establish internal procedures for its execution. Therefore, it’s essential to carry out analytical method validation in a way that aligns with both regulatory expectations and organisational standards.
I hope this article has helped elevate your understanding and equipped you with the knowledge needed to perform analytical method validation with confidence and precision.
Related:
Analytical method validation is the process of generating documented evidence that provides a high level of assurance that a specific process will consistently produce results meeting predefined specifications and quality attributes.
Analytical method validation tells about reliability of the Analytical method for intended use
Analytical Method Validation protocol is the manual of any Analytical validation work and it is Analytical method specific . It is written by the project coordinator and approved by the Analytical Research Manager, Quality Control Manager and Quality Assurance Manager
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