SOP for Analytical Method Validation

Purpose

To lay down a procedure for Analytical Method Validation of non-standard or modified methods for laboratory analysis.

Objective

To provide a documented procedure for Analytical Method Validation.

Scope

• To define role/responsibility of various persons responsible for Analytical Method Validation.
• This procedure is applicable to the analysis of samples using methodologies that are not established by the USP, BP, EP, JP, IP or any other Pharmacopoeia.

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Responsibility

• The Quality Assurance Manager is responsible for determining the level of validation required for each analytical method.
• Each analyst is responsible for the generation of data by the specific instructions of the Quality Assurance Manager/Quality Control Manager
• The analyst is also responsible for the initial review of the data package.
• The Quality Assurance Manager/Quality Control Manager is responsible for the secondary review of the data package to ensure that the analytical methodology used is validated by the instruction of this SOP.

Procedure

General Concepts

• Validation is the act of demonstrating and documenting a procedure that operates effectively.
• The discussion of the validation of analytical procedures is directed to the four most common types of analytical procedures:

1. Identification tests
2. Quantitative tests for impurities content
3. Limit tests for the control of impurities
4. Quantitative tests of the active moiety in samples of drug substance or drug product or other selected components in the drug product.

• Typical validation characteristics which should be considered are:

1. Accuracy
2. Precision
3. Specificity
4. Quantitation Limit
5. Linearity and Range
6. Robustness

Method Validation Parameter for the assay

Specificity:

Specificity is the ability to assess unequivocally the analyte in the presence of components that may be expected to be present. Typically these might include impurities, degradants,

matrix, etc.

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Linearity:

To be analyzed as per the proposed method. The results obtained is used to statistically evaluate the coefficient of determination (R2), standard error of estimate, and y-intercept.

Accuracy

The accuracy of the method is ascertained by the standard addition method at 3 levels. Standard quantity equivalent to 80%, 100% and 120% is to be added in the sample.

Precision

The precision of the chemical method is ascertained by carrying out the analysis as per the procedure and as per average weight taken for analysis. Repeat the analysis five times. Calculate the % assay, mean assay, % Deviation and % relative standard deviation and %RSD.

Range

The range of an analytical procedure is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy, and linearity.

Robustness

The robustness of an analytical procedure is a measure of its capacity to remain unaffected by small, but deliberate variations in method parameters and provides an indication of its reliability during normal usage.

Method Validation Parameter for Residual Solvent by GC

Specificity

Resolution of the analyte peak from the nearest peak: The solution of each of the analytes was injected separately and their retention time is noted. The standard working solution containing a mixture of the component being analyzed is also injected and each of the analyte peaks is checked for its resolution from the nearest.

Precision

Repeatability: Six replicate injections of standard solution for system precision should analyze as per the proposed method and from the chromatograms obtained the percentage % RSD is calculated.

Intermediate Precision: The purpose of this test is to demonstrate the intermediate precision of the method when the method is executed by a different analyst and on different day. The results obtained will be compared.

Linearity and Range

The solution of analyte solvent, having different concentrations should make separate from L.O.Q. concentration, which is 50% to 150%. The result obtained is statistically evaluated for the coefficient of determination (R2), standard error of estimate and y-intercept.

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LOD & LOQ

The limit of Detection (L.O.D.) was calculated as per below equation:

   

The limit of Quantification (L.O.Q.) was calculated as per the below equation:

Where, 

          σ = The standard deviation of the response from Linearity and range.

          S = The slope of the calibration curve Linearity & Range.

Accuracy / % Recovery (By Standard Addition Method)

• The accuracy of the method was ascertained by the standard addition method at 3 levels.
• Standard solution quantities equivalent to 50%, 100%, and 150% are added in the sample.
• The solution amount is analyzed by the proposed method and the chromatogram is obtained.
• The amount recovered by the method is compared to the amount added. Percent deviation is calculated at each level and a grand average across all the levels is also calculated.

          Methanol standard concentration –– 3000 ppm

          Acetic acid standard concentration –– 5000 ppm

          DMF standard concentration –– 880 ppm

Robustness

• The evaluation of robustness should be considered during the development phase and depends on the type of procedure under study. It should show the reliability of an analysis with respect to deliberate variations in method parameters.
• If measurements are susceptible to variation in analytical conditions, the analytical condition should be suitably controlled or a precautionary statement should be included in the procedure.
• One consequence of the robustness should be that a series of system suitability parameters (e.g. resolution test) is established to ensure that the validity of the analytical procedure is maintained whenever used.

Abbreviations

SOP: Standard Operating Procedure.

Annexure

Nil

Revision History

Nil

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