and analyzes reproducibility samples across different runs.

QC Reviewer: Reviews %CV calculations and verifies inter-run consistency.

QA Officer: Ensures reproducibility assessments are performed and documented for all applicable studies.

4. Accountability

The Bioanalytical Department Head is accountable for ensuring reproducibility assessments are planned, executed, and reviewed in compliance with regulatory requirements.

5. Procedure

5.1 Planning Reproducibility Assessment

1. Identify need for reproducibility assessment:
   • Multiple analytical runs within the same study
   • Samples analyzed on different days or instruments
   • Regulatory requirements such as for ISR (incurred sample reanalysis)
2. Select quality control (QC) samples or real subject samples previously analyzed in a qualifying run.
3. Prepare a Reproducibility Plan in Annexure-1: Reproducibility Assessment Plan.

5.2 Sample Selection and Reanalysis

1. Select at least 10% of total samples (preferably ISR samples) or a minimum of 20 samples, as per ICH M10 guidance.
2. Ensure selected samples:
   • Cover high, medium, and low concentration ranges
   • Have sufficient volume for repeat analysis
3. Reanalyze selected samples using the same validated method in a different run on a different day.

5.3 Data Calculation and Acceptance Criteria

1. Calculate % difference using:
   %Difference = ((Reanalysis - Original) / Mean) × 100
2. Acceptability criteria:
   • At least 67% of samples must show ≤20% difference from original result
3. Document all calculations in Annexure-2: Reproducibility Assessment Log.

5.4 Evaluation and Documentation

1. Summarize reproducibility results in the method validation or study report.
2. Attach reproducibility chromatograms, audit trails, and raw data for both runs.
3. Ensure all findings are signed and dated by the Analyst and Reviewed by QA.

5.5 Non-Conformance and Investigation

1. If < 67% of reanalyzed samples exceed 20% difference:
   • Initiate deviation report
   • Conduct root cause analysis (RCA)
   • Document CAPA using Annexure-3: Deviation and CAPA Form
2. Repeat assessment with new sample set, if justified.

6. Abbreviations

• BA/BE: Bioavailability/Bioequivalence
• ISR: Incurred Sample Reanalysis
• RCA: Root Cause Analysis
• CAPA: Corrective and Preventive Action
• QC: Quality Control
• %CV: Percent Coefficient of Variation

7. Documents

1. Reproducibility Assessment Plan – Annexure-1
2. Reproducibility Assessment Log – Annexure-2
3. Deviation and CAPA Form – Annexure-3

8. References

• ICH M10: Bioanalytical Method Validation
• FDA Guidance for Industry – Bioanalytical Method Validation (2022)
• WHO TRS No. 992 – Good practices for pharmaceutical quality control laboratories

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Reproducibility Assessment Plan

Study ID	Method Name	Analyst	Samples Selected	Run Date (Original/Repeat)
BAE-097	LC-MS/MS Method 3	Rajesh Kumar	PL-111 to PL-130	15/04/2025 / 17/04/2025

Annexure-2: Reproducibility Assessment Log

Sample ID	Original Result (ng/mL)	Reanalysis (ng/mL)	% Difference	Pass/Fail
PL-112	45.2	46.1	2.0%	Pass

Annexure-3: Deviation and CAPA Form

Deviation ID	Description	Root Cause	CAPA	Closure Date
DEV-2025-04	Failed reproducibility in ISR batch	Improper sample thawing	Re-training conducted	18/04/2025

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/01/2022	1.0	Initial SOP Release	New SOP	QA Head
17/04/2025	2.0	Added CAPA linkage and expanded annexures	Regulatory Alignment	QA Head