and quantified using the proposed method to validate its accuracy.

2. Scope

This SOP is applicable to all validation projects in the Analytical Method Development (AMD) department involving quantitative test methods such as assay, content uniformity, and impurity analysis in drug substances and drug products.

3. Responsibilities

• Validation Analyst: Performs spiking, sample preparation, and instrument analysis for recovery tests.
• Analytical Scientist: Designs the recovery study plan and interprets results.
• QA Officer: Reviews recovery data against acceptance criteria and approves the validation report.
• Head – AMD: Provides final authorization of accuracy study conclusions for regulatory submissions.

4. Accountability

The Head of AMD is accountable for ensuring that the accuracy and recovery study is statistically sound and conforms to ICH Q2(R2) requirements for method validation.

5. Procedure

5.1 Planning Recovery Levels

1. Select at least three concentration levels across the working range (e.g., 50%, 100%, and 150%).
2. Each level should be tested in triplicate (n=3) to ensure reproducibility.
3. Record planned concentrations in Annexure-1: Accuracy Level Planning Table.

5.2 Sample Preparation

1. Prepare placebo solution simulating the product matrix without API.
2. Spike known amounts of API standard into placebo at defined recovery levels.
3. Prepare test solutions as per the validated method, ensuring uniformity of dilution and mixing.
4. Prepare a standard solution for reference comparison.

5.3 Instrument Analysis

1. Inject spiked samples and standard solution into validated chromatographic or spectroscopic system.
2. Run system suitability prior to analysis.
3. Calculate recovery using the formula:
   
   Recovery (%) = (Measured Value / Theoretical Value) × 100
4. Record readings in Annexure-2: Recovery Data Sheet.

5.4 Acceptance Criteria

1. Recovery should be within:
   • 98.0% to 102.0% for assay methods
   • 80.0% to 120.0% for impurity or low-level tests
2. %RSD across replicates should be ≤2.0%.
3. All data must be statistically evaluated and tabulated.

5.5 Reporting

1. Summarize the study results in Annexure-3: Accuracy Summary Report.
2. Include individual recoveries, mean recovery, %RSD, and observations.
3. Attach chromatograms or spectral output for all samples analyzed.

6. Abbreviations

• API: Active Pharmaceutical Ingredient
• RSD: Relative Standard Deviation
• ICH: International Council for Harmonisation
• SOP: Standard Operating Procedure

7. Documents

1. Accuracy Level Planning Table – Annexure-1
2. Recovery Data Sheet – Annexure-2
3. Accuracy Summary Report – Annexure-3

8. References

• ICH Q2(R2) – Validation of Analytical Procedures
• FDA Guidance – Analytical Procedures and Methods Validation
• Internal Quality Manual – Method Validation Policy

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Accuracy Level Planning Table

Level	Theoretical Concentration (µg/mL)	Spike Volume	Matrix
50%	50.0	0.5 mL	Placebo
100%	100.0	1.0 mL	Placebo
150%	150.0	1.5 mL	Placebo

Annexure-2: Recovery Data Sheet

Level	Replicate	Measured Value (µg/mL)	Theoretical Value (µg/mL)	Recovery (%)
100%	1	99.4	100.0	99.4%
100%	2	100.2	100.0	100.2%
100%	3	98.9	100.0	98.9%

Annexure-3: Accuracy Summary Report

Level	Mean Recovery (%)	%RSD	Conclusion
100%	99.5%	0.67%	Pass

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
04/05/2025	2.0	Updated acceptance ranges and added annexure formats for consistency	Regulatory alignment with ICH Q2(R2)