1.0 OBJECTIVE
To lay down the procedure for the Analytical Method Validation
2.0 SCOPE
WHERE: This is the policy of Company that the written procedure to be followed for the validation of analytical method used for the analysis for the consistency of the method and must meet standards of accuracy and reliability of test protocols with predefined norms. WHEN : This analytical method validation is applicable at the time of Introduction of New Product and validation data not provided by tie up company. (Other than Pharmacopoeia products).
- Addition of new Raw material (other than Pharmacopoeias product).
- Ruggedness of pharmacopoeias drugs and drug product (if required). 3. RESPONSIBILITY
QC Executive/ QC Chemist.
4.0 Accountability:
4.1 Head – Quality Control shall be accountable for compliance of SOPs.
4.2 Head – Quality Assurance shall be accountable for compliance of SOPs.
5.0 PROCEDURE
5.1 Analytical Parameter (Definition and Procedure)
5.1.1 Linearity and Range
5.1.1.1 Definition of Linearity: Linearity of an analytical method is its ability to elicit test results that are
directly or by a well-defined mathematical transformation, proportional to the concentration of
Analyte in samples within a given range.
5.1.1.2 Definition of Range : The range of an analytical method is the interval between the upper and
lower level of analytic that have been demonstrated to be determined with a suitable level of
precision, accuracy and linearity using the method.
5.1.1.3 Procedure: The linearity should establish the relationship between the response of an analytic and
its concentration. For assay method linearity and range shall be done in the range of 80 to 120%
(For content uniformity 70 to 120%) of standard / sample preparation.
For related substances linearity and range should be done in the range of limit of Quantification and 120% of specification limits. A minimum of 5 data points is required in the range as specified above.
5.1.2 Precision
5.1.2.1 System Precision: The system precision is the agreement of five or more replicates that are obtained under identical conditions using some test methods.
5.1.2.2 Method Precision: The precision of an analytical method is the degree of agreement among individual test result when the method is applied repeatedly to multiple sampling of a homogeneous sample.
5.1.2.3 Procedure: The precision of an analytical method is by assaying a sufficient number of aliquots of a homogeneous sample to be able to calculate statistical estimates of standard deviation or relative standard deviation (coefficient of variation) of a series of measurement. Precision may be measured by the following three levels.
- Repeatability: It refers to use of the analytical procedure within a laboratory over a short
period time using the same analyst with of concern analytical method. The sample should be assessed using minimum of nine determinations covering the specified range for the procedure (i.e. three concentrations and three replicate of each concentration using minimum six determinations at 100 % of the test concentration).
Intermediate precision or ruggedness: The precision method on different days by different analyst and using different instruments. The sample shall prepare and analyzed as per the method, six times by two different analysts on different days and on different instruments.
- Reproducibility: Reproducibility refers to the use of the analytical procedure in different laboratories.
5.1.3 Accuracy
5.1.3.1 Definition: Accuracy of an analytical method is the closeness of test results obtained by that method to true values.
5.1.3.2 Procedure: For Active Pharmaceutical Ingredient (API), the accuracy is inferred from specificity, linearity, and precision data.
For drug product the accuracy shall be derived by performing, recovery experiments by spiking API in the placebo in triplicate at 80% to 120% of assay sample preparation. For Related Substances (API and Drug Product) the accuracy shall be shown by performing recovery experiment by spiking known impurities in the same in triplicate at 50, 100, and 120% of specifications.
5.1.4 Specificity
5.1.4.1 Definition: The specificity is defined as the ability to assess clearly the analyte, in the presence of components that may be expected to be present such as impurities, degradation product and matrix components.
5.1.4.2 Procedure:
Assay Method: In case of assay demonstration of specification requires that it can be shown that the procedure is unaffected by presence of impurities/related substances or excipient with the analytic peak. In practice this can be done by spiking the drugs substances with appropriate level of impurity (1%) or excipient in triplicate and demonstrating that the assay result is unaffected by the presence of these extraneous materials.
Related Substances method: This is done by spiking all known impurities at 1% level and showing the separation of all impurities from the main peak and from each other and also demonstrating peak purity.
5.1.5 Limit of detection
5.1.5.1 Definition: It is the lowest amount of analyte in a sample that can be detected but not necessary to be quantified.
5.1.5.2 Procedure: For non-instrumental method the detection limit is generally determined by the analysis of sample with known concentration of analyte and by establishing the minimum level at which the analyte can be reliably detected.In case of instrumental analytical procedures measure the magnitude of analytical back ground response by analysis number of analyte samples of different concentrations at detection level (1: 2 or 3 signal of noise ratio) and calculating limit of detection by following method.
LOD = 3.3 × S.E.
Slop
Where S.E. (Standard Error) and slop are calculated by number of concentration at LOD level.
5.1.6 Limit of Quantification
Definition: It is the lowest amount of analyte in a sample at which that can be determined with acceptable precision and accuracy under the stated experimental conditions. 5 to 10% precision is acceptable.
Procedure: For non-instrumental method the Quantification limit is generally determined by the analysis of sample with known concentration of analyte and by establishing the minimum level at which the analyte can be Quantify reliably.
In case of instrumental analytical procedures measure the magnitude of analytical background response by analysis number of analyte samples of different concentrations at Quantification level (1: 10 or 20 signal of noise ratio) and calculating limit of Quantification by following method.
LOQ = 10 × S.E.
Slop
Where S.E. (Standard Error) and slop are calculated by number of concentration at LOQ level.
5.1.7 Robustness
5.1.7.1 Definition: Robustness is to show the ability of method to remain unaffected by small but deliberate variations in the method parameters.
5.1.7.2 Procedure: The robustness of method shall be done by following deliberately change in the method.
The stability of analyte in solution should be demonstrated to show the robustness of the method.
- i) Change in flow rate ±10%.
- ii) Change in pH buffer/mobile phase by ± 0.2 unit.
iii) Change in organic content of mobile phase by ± 2%. iv) Change in column temperature by ± 2%.
5.2 VALIDATION PLAN
5.2.1 Following analytical parameter shall be validated before implementation of analytical method.
- Assay
- Determination of related substances.
- Dissolution Rate tests.
5.2.2 If the assay procedure for content uniformity is different from the composite assay the test method shall be validated.
5.2.3 In case of HPLC method, the system suitability shall be established before the analysis.
5.2.4 For pharmacopoeia drugs and drugs products only robustness parameter shall be validated.
5.2.5 For non-pharmacopoeia drugs and drug product, the method shall be validated for the following parameters.
5.2.6 Raw Material
- Assay
- i) Linearity & range ii) Precision – System precision, Method precision iii)Robustness iv) Specificity v) Accuracy
- Related substances
- i) Linearity ii) Precision – System Precision, Method Precision iii) Specificity iv) Robustness v) Limit of detection vi) Limit of quantification vii) Accuracy
5.2.7 Finished Product
Assay of active material(s) and preservative(s) (if any)
- i) Linearity ii) Precision – System Precision
Method Precision
iii) Accuracy
iv) Robustness
v) Specificity
5.2.8 Dissolution Rate
- i) Linearity ii) Precision – System Precision
Method Precision
iii) Robustness
Dissolution rate for time release tablet
i) Linearity (from 0 to 120 %. of active drug)
ii) Time stability
iii) Precision – System Precision
Method Precision
iv) Robustness
5.3 Revalidation
A partial or complete revalidation of the analytical method shall be done according to following defined change and revalidation criteria.
| S.No. | CHANGE | PARAMETER TO BE VALIDATED | |
| i | Change in composition of drugs | :
: : |
Specifications
Linearity Precision |
| ii | Revision in specification | :
: |
Linearity
Accuracy |
| iii | Change in Master Formula (Excipients) | :
: : |
Specifications
Linearity Precision |
| iv | Change in analytical Procedure | : | Complete validation |
5.4 Acceptance Criteria
The acceptance criteria for the analytical method validation will be followed:
| Characteristics | Acceptance Criteria | |
| Linearity | : | R2 0.99, similar response ratio |
| Precision – System | : | RSD < 2% |
| Precision – Method | : | RSD < 2% |
| Precision – Repeatability / Reproducibility | : | % R & R <20% |
| Accuracy | : | 98 to 102 % (in range 50 to 150%) |
| Specificity | : | No interference |
| Quantification | : | Signal noise ratio 10:1 or 10 X S.dev / Slop |
| Detection | : | Signal noise ratio 2:1 or 3.3 X S.dev / Slop |
| Range | : | Concentration where data can be reliably determine (98 t0 102 % recovery) |
| Robustness | : | Recovery 98 t0 102 % |
6.0 REFERENCE(s)
6.1 WHO Guidelines
7.0 FORMAT(s)
7.1 Annexure-I Analytical Method Validation Log book ,Format No.
8.0 ABBREVIATION(s)
8.1 SOP : Standard Operating Procedure
8.2 Dept. : Department
8.3 No. : Number
8.4 QA : Quality Assurance
8.5 cGMP : current Good Manufacturing Practices
8.6 NMT : Not More than
9.0 Distribution
9.1 Controlled Copy No. QC/01 : Head of Department – Quality Control
9.2 Master Copy : Head – QUALITY ASSURANCE
- TRAINING:
10.1 Trainer — Head – Quality Control
10.2 Trainee — Q.C Chemist.
10.3 Period — 30 minutes
11.0 REASON FOR REVISION
11.1 Periodic revision
