analytical method validation checklist

Define the purpose and scope of the method.

Identify the analytes of interest.

Select the appropriate analytical technique.

Optimize method conditions (e.g., temperature, pH, solvents).

Conduct preliminary experiments for feasibility.

Assess the method's ability to differentiate between analytes and potential interferences.

Evaluate the impact of matrix components on analyte detection.

Perform experiments to identify any potential cross-reactivity.

Establish a calibration curve over an appropriate concentration range.

Determine the correlation coefficient (R²) of the calibration data.

Evaluate the linearity by performing statistical analysis (e.g., ANOVA).

Perform recovery studies by spiking known concentrations of analytes into the matrix.

Calculate percentage recovery and assess consistency across the range.

Compare results with a reference method if available.

Conduct repeatability tests (intra-day precision) by analyzing multiple replicates.

Assess intermediate precision (inter-day precision) by performing tests on different days.

Calculate relative standard deviation (RSD) for both repeatability and intermediate precision.

Determine the limit of detection (LOD) and limit of quantitation (LOQ).

Use signal-to-noise ratio or standard deviation of the blank for calculations.

Validate LOD and LOQ through practical experiments.

Assess the method's ability to remain unaffected by small variations in method parameters (e.g., temperature, time, pH).

Conduct experiments to identify critical parameters and their impact on the method.

Document findings and determine acceptable ranges for parameters.

Evaluate the stability of the analytes in the matrix over time.

Assess the stability of the prepared samples under various storage conditions.

Conduct freeze-thaw studies if applicable.

Maintain comprehensive records of all validation studies.

Document all experimental conditions, results, and statistical analyses.

Prepare a validation report summarizing all findings and conclusions.

Ensure adherence to relevant guidelines (e.g., ICH, FDA, EMA).

Review validation protocols and reports for compliance with regulatory standards.

Prepare for potential audits by regulatory bodies.