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.