Objective:To lay down the procedure for Analytical Method Validation.
Scope:This SOP is applicable for all the products or whenever changes in formulation & specification of the products in quality control department.
Responsibility: Officer or above of QC Laboratory shall be responsible for follow the procedure mentioned in this SOP.
Accountability:Department Head & QA Head shall be accountable for implementation of this SOP
Abbreviations and Definitions
LOQ: Limit of Quantification
LOD: Limit of Detection
HPLC:High Performace Liquid Chromatography
AMV:Analytical Method Validation
All the instruments used in AMV must be calibrated.
Analyst qualification must be ensured who performing the AMV.
All the measuring glassware used in AMV must be calibrated or class ‘A’ type.
Element Data required for validation Annexure- I
Analytical method validation is performed for tests like Assay, Dissolution, Impurities/ Related Substances individually.
In case of Assay or Impurities-profile, system suitability is essential prior to start the validation activity.
Any analytical method shall be revised through change control procedure and shall be duly approved by Quality Assurance and then it undergoes for validation.
Whenever there is a need to change critical test parameters,the method is re-validated with the revised parameters & conditions.
Following Analytical Performance Characteristic shall be followed for Analytical method validation.
Specificity is the ability to assess unequivocally the analyte, in the presence of components that may be expected to be present, such as impurities, degradation products, and matrix components
Ruggedness (Intermediate Precision):
Intermediate precision (ruggedness) is the precision obtained when the assay is performed by multiple analysts, using multiple instruments, on multiple days, in one laboratory. Different sources of reagents and multiple lots of columns are included in this study. Intermediate precision results are used to identify which of the above factors contribute significant variability to the final result. Depending on time and resources, the method can be tested on multiple days, analysts, instruments, etc.
Precision is the measure of how close the data values to each other for a number of measurements under the same analytical conditions.
Accuracy is the exactness of an analytical method or the closeness of agreement between the value (which is accepted either as a conventional true value or an accepted reference value and the value found. It is measure as the percent of analyte recovered from matrix components.
For the assay of the drug substance, accuracy measurements are obtained by comparison of the results with the analysis of a standard reference material or by comparison to a second, well-characterized method.
For the assay of the drug product, accuracy is evaluated by analyzing synthetic mixtures spiked with known quantities of components.
For the quantization of impurities, accuracy is determined by analyzing sample (drug substance or drug product) spiked with known amounts of impurities.
Linearity & Range:
The 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.
The range of an analytical method is the interval between the upper and lower levels of analyte (including these levels) that have been demonstrated to be determined with a suitable level of precision, accuracy and linearity using the method as written. In practice, the range is determined using the data from the linearity of response and accuracy studies.
Stability of Solution:
The period of time a solution can be held before analysis without compromising accuracy.This delay is beyond that included in the method procedure and anticipates unexpected instrumental delays.
Robustness is the capacity of a method to remain unaffected by small, deliberate variations in method parameters and this can be evaluated by varying method parameters such as flow rate, pH of buffer/mobile phase, composition of mobile phase, temperature, wavelength etc., within a realistic range and the quantitative influence of the variables is determines. If the influence of the parameter is within a previously specified tolerance, the parameter is said to be within the method’s robustness range.
Analyte loss is commonly encountered via membrane filter adsorption of acidic, basic, and neutral compounds of pharmaceutical interest. Acidic compounds exhibit the greatest adsorption losses, basic compounds exhibit an intermediate amount, and neutral compounds exhibit least losses. The primary objective of this study is to determine the extent of membrane filter adsorption of analyte(s) during filtration of test solution through membrane filter as per test method.
Limit of Detection (LOD):
The limit of detection is the point at which a measured value is larger than the uncertainty associated with it. It is the lowest concentration of analyte in a sample that can be detected but not necessarily quantified.
Limit of Quantification (LOQ):
The limit of quantification is the lowest level of analyte that can be accurately and precisely measured. This limit is required only for impurity methods and is determined by reducing the analyte concentration until a level is reached where the precision of the method is unacceptable.
Revalidation of Analytical Method:
Validation procedure should be carried out prior to implementing the said procedure for analysis. It should be repeated whenever there is any change in formulation, specification or composition of the product takes place & any change in analytical method.
Acceptance criteria are followed as mentioned in individual validation protocol for particular product
Forms and Records (Annexures)
Data element required for validation -Annexure-I
Master copy – Quality Assurance
Controlled copies – Quality Assurance & Quality Control
Reason for Revision
Data element required for validation
|Analytical Performance Characteristics||Category I||Category II||Category III||Category IV|
Note: * May be required, depending on the nature of the specific test.
Category I : Analytical methods for quantitation of major components of bulk drug substances or active ingredients (including preservatives in finished pharmaceutical products.
Category II : Analytical methods for determination of impurities in bulk drug substances or degradation compounds in finished pharmaceutical products. These methods include quantitative assays and limit test.
Category III : Analytical methods for determination of performance characteristic ( e.g., dissolution, drug release).
Category IV : Identification tests.