loss or interference.

2. Scope

This procedure applies to Analytical Method Development (AMD) for microbiological testing, quantitative chemical analyses, and impurity profiling where sample recovery impacts result interpretation and method validation.

3. Responsibilities

• Analytical Scientist: Plans and executes the recovery evaluation as per protocol.
• Microbiologist: Performs microbial recovery studies and ensures strain viability and proper dilution.
• QA Officer: Reviews all recovery records for accuracy, completeness, and regulatory alignment.
• Head – AMD: Approves recovery protocols and interprets outcomes in relation to method robustness.

4. Accountability

The Head of Analytical Method Development is accountable for the scientific reliability and compliance of recovery evaluation protocols and data generated.

5. Procedure

5.1 Recovery Study Design

1. Define target component: drug substance, degradation product, microbial organism, etc.
2. Select appropriate matrix/sample (e.g., placebo, blank, water, API, formulation).
3. Choose test concentration levels: low, medium, and high range within method quantification span.
4. Perform triplicate spiking at each level to assess precision and recovery consistency.
5. Record recovery plan in Annexure-1: Recovery Study Design Form.

5.2 Preparation of Spiked Samples

1. Prepare known standard stock solution or microbial inoculum of validated concentration.
2. Accurately spike into blank matrix to mimic sample behavior.
3. Perform parallel analysis of unspiked (blank) and standard/reference solutions.
4. Label all samples clearly and document volumes and concentrations.
5. Log details in Annexure-2: Spiking and Sample Preparation Log.

5.3 Analytical Measurement

1. Analyze spiked samples using the method under development or validation.
2. Use the same instrument settings, analyst, and run conditions as intended for routine use.
3. Calculate recovery using the formula:
   
   Recovery (%) = (Measured Amount / Theoretical Amount) × 100
4. Document raw data and calculations in Annexure-3: Analytical Data Sheet.

5.4 Acceptance Criteria

1. Acceptance range depends on test type:
   • Microbial: 50%–200% recovery of spiked organisms
   • Chemical: 98%–102% (assay); 95%–105% (impurities)
2. If recovery falls outside limits, investigate dilution error, matrix interference, or analyst technique.
3. Repeat study if deviation persists or modify method accordingly.

5.5 Reporting and Review

1. Summarize results in a tabular format showing:
   • Sample ID
   • Spiked concentration
   • Recovered amount
   • % Recovery
2. Highlight any outliers or trends observed across different concentration levels.
3. Attach chromatograms or microbial plate photos (if applicable).
4. Complete Annexure-4: Recovery Summary Report.

6. Abbreviations

• AMD: Analytical Method Development
• CFU: Colony Forming Unit
• SOP: Standard Operating Procedure
• LOD: Limit of Detection
• LOQ: Limit of Quantification

7. Documents

1. Recovery Study Design Form – Annexure-1
2. Spiking and Sample Preparation Log – Annexure-2
3. Analytical Data Sheet – Annexure-3
4. Recovery Summary Report – Annexure-4

8. References

• ICH Q2(R1) – Validation of Analytical Procedures
• USP General Chapter <1225> – Validation of Compendial Procedures
• ICH Q6A – Specifications: Test Procedures and Acceptance Criteria

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Recovery Study Design Form

Analyte	Matrix	Spiking Levels	Planned By
Paracetamol	Syrup Base	80%, 100%, 120%	Rajesh Kumar

Annexure-2: Spiking and Sample Preparation Log

Sample ID	Volume	Spiked Conc.	Prep Date	By
REC-128-01	10 mL	100 µg/mL	18/05/2025	Sunita Reddy

Annexure-3: Analytical Data Sheet

Sample	Theoretical Amount	Measured Amount	% Recovery
REC-128-01	100 µg	98.2 µg	98.2%

Annexure-4: Recovery Summary Report

The method demonstrated recovery rates within 98%–101% across three concentration levels. No matrix interference observed. Method suitable for validation and routine analysis.

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
04/05/2025	2.0	Expanded to include microbial recovery scope and ICH Q6A references	Annual SOP Review