the true values by spiking known quantities of the analyte into the matrix.

2. Scope

This SOP applies to all analytical methods being developed and validated within the Analytical Method Development (AMD) department for pharmaceutical drug substances and drug products where quantitative analysis is required.

3. Responsibilities

• Analytical Scientist: Responsible for planning and executing the accuracy study and documenting results.
• QA Reviewer: Reviews accuracy study data and verifies statistical validity and protocol compliance.
• Group Lead: Approves accuracy results and ensures compliance with validation requirements.

4. Accountability

The AMD Section Head is accountable for ensuring that all accuracy studies are performed as per ICH Q2(R1) guidelines and documented in a compliant and reproducible manner.

5. Procedure

5.1 Planning the Accuracy Study

1. Identify the appropriate matrix (e.g., placebo, diluent) to which the analyte will be spiked.
2. Select at least three concentration levels: typically 80%, 100%, and 120% of the test concentration.
3. Each level should be prepared in triplicate to evaluate reproducibility.

5.2 Preparation of Accuracy Samples

1. Weigh appropriate quantities of analyte standard using calibrated analytical balance (Annexure-1).
2. Spike the analyte into placebo or matrix in volumetric flasks to achieve target concentrations (Annexure-2).
3. Ensure homogeneous mixing by vortexing or mild sonication as appropriate.
4. Prepare blank placebo and matrix controls as part of the set.

5.3 Sample Analysis

1. Analyze the accuracy samples using the validated analytical procedure (HPLC, UV, etc.).
2. Inject all samples in a single run with appropriate system suitability checks.
3. Document retention time, area, and any system suitability parameters in the run log.

5.4 Calculation of Recovery

1. Calculate the percent recovery at each level using the formula:
   % Recovery = (Measured Value / Added Value) × 100
2. Evaluate the mean, standard deviation (SD), and % Relative Standard Deviation (RSD) for each level.
3. Acceptable recovery: 98–102% for assays and 80–120% for impurities depending on regulatory expectations.

5.5 Data Evaluation and Acceptance Criteria

1. Review all raw chromatograms, calculations, and recovery data.
2. If any sample falls outside the acceptance range, perform root cause analysis.
3. Document any repeat analysis, justification, and final approved data set.

5.6 Documentation and Reporting

1. Complete Annexure-3 Accuracy Data Summary Table.
2. Attach all analytical raw data, sample preparation sheets, chromatograms, and electronic data output.
3. Review and approve the study report as part of the final method validation report.

6. Abbreviations

• SD: Standard Deviation
• RSD: Relative Standard Deviation
• HPLC: High Performance Liquid Chromatography
• ICH: International Council for Harmonisation

7. Documents

1. Annexure-1: Standard Weight Log
2. Annexure-2: Accuracy Sample Preparation Sheet
3. Annexure-3: Accuracy Recovery Table

8. References

• ICH Q2(R1): Validation of Analytical Procedures
• USP General Chapter <1225>: Validation of Compendial Procedures
• FDA Guidance on Analytical Procedures and Methods Validation

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Ashutosh Sharma	Meenal Shah	Dr. Manish Rathi
Designation	Method Development Chemist	QA Executive	Head – AMD
Department	Analytical Method Development	Quality Assurance	Analytical Method Development

11. Annexures

Annexure-1: Standard Weight Log

Standard Name	Weight Taken (mg)	Batch No.	Prepared By
API-A	25.0	ST/2025/041	A. Sharma

Annexure-2: Accuracy Sample Preparation Sheet

Level	Target Concentration (µg/mL)	Actual Spiked	Matrix	Remarks
80%	40	40.2	Placebo-A	Within acceptable range
100%	50	49.8	Placebo-A	Pass
120%	60	59.9	Placebo-A	Pass

Annexure-3: Accuracy Recovery Table

Level	% Recovery (Replicate 1)	Replicate 2	Replicate 3	Average	% RSD
80%	99.5	98.9	100.1	99.5	0.6%
100%	99.2	99.7	100.0	99.6	0.4%
120%	101.0	100.6	100.8	100.8	0.2%

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/06/2025	2.0	Expanded acceptance criteria and clarified statistical evaluation	Annual Review	Dr. Manish Rathi
12/07/2022	1.0	Initial SOP Release	New SOP	QA Head