analysis of plasma samples using LC-MS/MS instrumentation.

3. Responsibilities

• Analyst: Reviews system suitability, evaluates calibration and QC performance, and documents results.
• Reviewer/Supervisor: Verifies correctness of curve parameters, QC recovery, and overall run validity.
• QA: Performs audits on run acceptance decisions and verifies compliance with SOP and validation protocols.

4. Accountability

The Head of Bioanalytical Laboratory is accountable for ensuring that only runs meeting the acceptance criteria are used for reporting pharmacokinetic data.

5. Procedure

5.1 Calibration Curve Review

1. Each run must include at least six non-zero calibration standards covering the validated range.
2. Plot peak area ratio (analyte/internal standard) against nominal concentration.
3. Evaluate curve using appropriate regression (linear with 1/x² weighting preferred unless otherwise validated).
4. Acceptance Criteria:
   • ≥75% of non-zero calibrators should be within ±15% of nominal (±20% for LLOQ).
   • Correlation coefficient (r) should be ≥ 0.99.
5. Document the regression equation, r² value, and residuals in Annexure-1: Calibration Curve Summary Log.

5.2 QC Sample Review

1. Each batch must contain QC samples at three levels (LQC, MQC, HQC) in duplicate or more.
2. Acceptance Criteria:
   • At least 67% of QCs should be within ±15% of nominal concentration.
   • At least 50% of QCs at each level must meet the above criteria.
3. Calculate accuracy and precision for each QC level and document in Annexure-2: QC Acceptance Summary.
4. If criteria are not met, consider:
   • Repeating the run
   • Investigating cause of deviation
   • Documenting result as a failed run

5.3 System Suitability Checks

1. Confirm that:
   • Retention times are within ±2% of validated range
   • Peak shape is consistent
   • Blank samples show no carryover
2. Document performance in Annexure-3: System Suitability Log.

5.4 Documentation and Approval

1. All results must be reviewed by the supervisor and QA prior to data reporting.
2. Approved curves and QCs are stored in electronic and physical format.
3. Any deviation from acceptance criteria must be logged in Annexure-4: Deviation Justification Record.

6. Abbreviations

• BA/BE: Bioavailability/Bioequivalence
• QC: Quality Control
• LLOQ: Lower Limit of Quantification
• LC-MS/MS: Liquid Chromatography with Tandem Mass Spectrometry

7. Documents

1. Calibration Curve Summary Log – Annexure-1
2. QC Acceptance Summary – Annexure-2
3. System Suitability Log – Annexure-3
4. Deviation Justification Record – Annexure-4

8. References

• ICH M10: Bioanalytical Method Validation
• US FDA Guidance for Industry: Bioanalytical Method Validation (2022)
• Sponsor Bioanalytical Method Validation Protocol

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Calibration Curve Summary Log

Batch ID	Equation	r² Value	Acceptance	Reviewer
BE1278-02	y = 0.0043x + 0.002	0.9987	Accepted	Sunita Reddy

Annexure-2: QC Acceptance Summary

QC Level	Replicate No.	Measured Value	Nominal	% Difference	Acceptance
LQC	1	18.5	20.0	-7.5%	Accepted

Annexure-3: System Suitability Log

Run Date	Retention Time (RT)	RT Acceptance	Carryover	Reviewer
17/04/2025	3.45	Yes	No	Rajesh Kumar

Annexure-4: Deviation Justification Record

Date	Batch	Issue	Justification	Approved By
17/04/2025	BE1278-03	One QC failed	Reinjected and verified	QA Manager

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/01/2022	1.0	Initial version	New SOP	QA Head
17/04/2025	2.0	Added deviation documentation and system suitability logging	Regulatory update	QA Head