What is the Method Validation Steps According to ICH and USP Guidelines? and What is Acceptance Criteria for this Steps ?

Method Validation:

“The process by which it is established, through laboratory studies, that the performance

characteristics of the method meet the requirements for its intended purpose”

Specificity:

“Specificity is the ability to measure accurately and specifically the analyte of interest in the

presence of other components that may be expected to be present in the matrix.

Selectivity:

“Selectivity is a relative term to describe to which extent particular analytes in mixtures or

matrices can be measured without interferences from other components of similar behaviour

Linearity:

” The ability of the method to elicit test results that are directly, or by a well-defined

mathematical transformation, proportional to analyte concentration within a given range.”

Precision:

Precision the dgree of agreement among individual test result when an analytical method is

used repeatedly to multiple samplings of homogeneous sample

a) Intraday Precision in different time but in the same day: to ensure that the methods are stabile and repeatable in same day but long time

b) Intraday Precision Repeatability (in the same time) : the replicate test in short time under the same condition

c) Interday Precision: Intermediate Precision:-

to ensure that the method is strong in the real routine analysis by different days

Accuracy:

“The closeness of test results to the true value”

Robustness :

The capacity of a method to remain unaffected by small, deliberate variations in method

parameters ; a measure of the reliability of a method, often used to set the system suitability

parameters of a method.

System Suitability :

to ensure that the validation method had a suitable system during validation test by

calculated Tailing factor ,Theoretical plate count (N) …

Method Validation Acceptance Criteria According to ICH and USP Guidelines

1. Linearity

ICH/USP Criteria:

Correlation coefficient (R²) ≥ 0.995.

Linear regression equation: y=mx+b.

2. Accuracy (Recovery %)

ICH/USP Criteria: Recovery percentage should be between 98% and 102%.

3. Precision

ICH/USP Criteria:

RSD ≤ 2% for intraday precision.

RSD ≤ 3% for interday precision.

4. Sensitivity (LOD and LOQ)

ICH/USP Criteria:

LOD (Limit of Detection): Lowest concentration detectable.

LOQ (Limit of Quantification): Lowest concentration measurable with acceptable precision.

5. Robustness

ICH/USP Criteria:

RSD ≤ 3% under minor variations in operating conditions (e.g., flow rate or pH).

6. System Suitability

ICH/USP Criteria:

Number of theoretical plates (N) > 2000.

Resolution (Rs) > 1.5.

Aug 18

Mohammad Asaad Mousa,

Kindly refer following Guidelines

1) VALIDATION OF ANALYTICAL PROCEDURES Q2(R2).

2) Validation of Chromatographic Methods" (CDER, 1994)

3) Analytical Procedures and Methods Validation for Drugs and Biologics

U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) Center for Biologics Evaluation and Research (CBER) July 2015 Pharmaceutical Quality/CMC

The specific steps and acceptance criteria are determined by the test's purpose, but general parameters are evaluated in every validation. Both the International Council for Harmonisation (ICH) and the United States Pharmacopeia (USP) provide guidelines for validating analytical methods, which are crucial for ensuring data reliability in the pharmaceutical industry

Method validation steps (ICH Q2(R1) and USP )

The following steps outline the key parameters that must be validated to confirm a method's suitability for its intended purpose.

1. Specificity

• Definition: The ability to measure the analyte accurately and without interference from other components that might be present in the sample, such as impurities, degradants, and excipients.
• How to test: Challenge the method by analyzing samples containing known impurities or potential interfering substances and verify that they do not affect the analyte's measurement. For stability-indicating methods, forced degradation studies are performed to ensure the method can separate the active ingredient from its degradation products.
• Acceptance criteria:The analyte peak must be well-separated from all other peaks. For identification tests, the test must confirm the identity of the analyte. For impurity tests, the method must separate the analyte from all potential impurities.

2. Accuracy

• Definition: The closeness of the measured value to the true value. It is often expressed as the percentage of recovery.
• How to test: Use a fully validated reference method or spike a placebo sample with a known amount of analyte to demonstrate consistent recovery across the method's range.
• Acceptance criteria: For assays, the typical acceptance range for recovery is 98–102%, though the exact criteria should be justified based on the analyte's concentration and complexity.

3. Precision

• Definition: The degree of agreement among individual test results when the method is repeatedly applied to multiple samples. It is evaluated at three levels:Repeatability: Precision under the same operating conditions over a short time interval. Intermediate precision: Precision within a laboratory over different days, with different analysts, or using different equipment. Reproducibility: Precision between different laboratories, typically relevant for standardization.
• How to test: Analyze multiple aliquots of a homogeneous sample. This is often done by preparing and testing at least six replicates at 100% concentration for repeatability testing.
• Acceptance criteria: Typically expressed as the relative standard deviation (RSD). For assays, a repeatability RSD of ≤ 1% to 2% is common, while for impurity determinations, a higher RSD (e.g., up to 10% to 15%) may be acceptable, particularly near the quantitation limit.

4. Linearity and Range

• Definition:Linearity: The method's ability to produce results that are directly proportional to the concentration of the analyte within a given range. Range: The interval over which the method consistently demonstrates acceptable accuracy, precision, and linearity.
• How to test: Prepare and analyze a series of at least five standard solutions across the concentration range to be validated. Plot the analyte response versus concentration and perform a statistical analysis, such as linear regression.
• ) for the regression line must be high, typically ≥ 0.995 for most chromatographic methods. The range is confirmed to cover the required operating concentration. For example, a range of 80–120% of the target concentration is standard for assays.Acceptance criteria: The correlation coefficient (𝑟2📷r2r squared

5. Detection Limit (LOD) and Quantitation Limit (LOQ)

• Definition: LOD is the lowest detectable analyte amount, while LOQ is the lowest quantifiable amount with acceptable accuracy and precision.
• How to test: These can be determined using signal-to-noise ratios or calculations based on the standard deviation of the response and calibration curve slope.
• Acceptance criteria: A signal-to-noise ratio of at least 3:1 is typical for LOD, and at least 10:1 for LOQ. These are particularly important for impurity and limit tests.

6. Robustness

• Definition: The method's reliability when subjected to minor variations in parameters.
• How to test: Introduce small changes to parameters (e.g., pH, flow rate) and check the impact on results.
• Acceptance criteria: Results should not be significantly affected by these variations. An RSD typically ≤ 3% is often expected.

Thank you Mohammad for the recommendation. You are a scholar and a gentleman.