spiking known quantities into the sample matrix.

2. Scope

This procedure is applicable to all new or modified analytical methods intended for the quantification of active pharmaceutical ingredients (APIs), excipients, and finished drug products at various concentration levels in the Analytical Method Development department.

3. Responsibilities

• Analyst: Prepares and analyzes spiked samples at specified levels and records all data and observations.
• Method Development Scientist: Reviews data and validates calculations, checks compliance with acceptance criteria.
• QA Representative: Audits raw data, recovery calculations, and ensures adherence to protocol and SOP.

4. Accountability

The Head of Analytical Method Development is accountable for ensuring that accuracy studies are properly conducted, documented, reviewed, and retained in accordance with regulatory standards including ICH Q2(R1) and applicable GMP guidelines.

5. Procedure

5.1 Selection of Levels

1. Choose at least three concentration levels representing the lower (80%), target (100%), and upper (120%) range of the analytical method.
2. Prepare each level in triplicate using the same matrix as the intended sample (e.g., placebo or formulation blend).

5.2 Preparation of Recovery Samples

1. Prepare the placebo matrix that mimics the sample composition, excluding the active ingredient.
2. Spike the placebo with known amounts of standard solution to achieve 80%, 100%, and 120% of target concentration.
3. Process the spiked samples as per the test method, including any extraction or filtration steps.

5.3 Analysis

1. Inject each prepared recovery sample into the analytical instrument under validated chromatographic or spectroscopic conditions.
2. Calculate the amount recovered using the standard calibration curve or direct comparison to standard solutions.
3. Determine % recovery using the formula: % Recovery = (Recovered amount / Added amount) × 100.

5.4 Data Compilation and Interpretation

1. Calculate the mean % recovery and % Relative Standard Deviation (%RSD) for each level.
2. The acceptance criteria for accuracy are generally 98.0–102.0% for assay methods. Tighter or wider limits may be used based on product requirements and method sensitivity.
3. Compile results using Annexure-1 and summarize in Annexure-2: Accuracy Summary Sheet.

5.5 Reporting

1. Prepare a report containing methodology, sample preparation, recovery data, graphs (if applicable), and conclusion.
2. Submit the report to the QA department for review and approval.

6. Abbreviations

• API: Active Pharmaceutical Ingredient
• RSD: Relative Standard Deviation
• ICH: International Council for Harmonisation
• QA: Quality Assurance

7. Documents

1. Annexure-1: Accuracy Recovery Data Sheet
2. Annexure-2: Accuracy Summary Sheet
3. Analytical Method Validation Protocol

8. References

• ICH Q2(R1): Validation of Analytical Procedures
• USP Chapter <1225> Validation of Compendial Procedures
• 21 CFR Part 211

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Vivek Rawat	Sneha Joshi	Dr. Arun Patel
Designation	Analyst	QA Reviewer	Head – AMD
Department	Analytical Method Development	Quality Assurance	Analytical Method Development

11. Annexures

Annexure-1: Accuracy Recovery Data Sheet

Level	Added (mg)	Recovered (mg)	% Recovery
80%	8.0	7.92	99.0%
80%	8.0	8.04	100.5%
80%	8.0	7.96	99.5%
100%	10.0	10.1	101.0%
100%	10.0	9.9	99.0%
100%	10.0	10.0	100.0%
120%	12.0	12.1	100.8%
120%	12.0	11.8	98.3%
120%	12.0	12.2	101.6%

Annexure-2: Accuracy Summary Sheet

• 80% Level: Mean Recovery = 99.7%, RSD = 0.76%
• 100% Level: Mean Recovery = 100.0%, RSD = 1.0%
• 120% Level: Mean Recovery = 100.23%, RSD = 1.69%

Conclusion: The analytical method demonstrates acceptable accuracy across the tested range.

Revision History

Revision Date	Revision No.	Description	Reason	Approved By
01/06/2025	2.0	Expanded acceptance criteria and updated table formats	Annual SOP Review	Dr. Arun Patel
15/07/2022	1.0	Initial SOP Release	New Method Validation	QA Head