Standard Operating Procedure for Stability Testing of Analyte in Plasma for BA/BE Studies
| Department | BA-BE Studies |
|---|---|
| SOP No. | SOP/BA-BE/173/2025 |
| Supersedes | SOP/BA-BE/173/2022 |
| Page No. | Page 1 of 10 |
| Issue Date | 17/04/2025 |
| Effective Date | 20/04/2025 |
| Review Date | 17/04/2026 |
1. Purpose
To outline the procedure for evaluating the stability of analytes in plasma samples under various conditions to ensure the integrity of study samples analyzed during bioavailability/bioequivalence (BA/BE) studies.
2. Scope
This SOP applies to all stability tests conducted on plasma samples for validating storage and handling conditions during method validation and sample analysis phases of BA/BE studies.
3. Responsibilities
- Analyst: Performs stability experiments as per protocol and records results.
- Reviewer/Supervisor: Verifies stability data and compliance with acceptance criteria.
- QA: Reviews documentation and ensures regulatory adherence.
4. Accountability
The Bioanalytical Lab Manager is accountable for ensuring that plasma sample stability data is valid, reproducible, and documented in accordance with ICH M10 and regulatory expectations.
5. Procedure
5.1 Types of Stability Tests
- Short-Term Stability: Room temperature exposure for 4-24 hours.
- Long-Term Stability: Storage at -20°C or -70°C for the validated duration (e.g., 30, 60, 90 days).
- Freeze-Thaw Stability: Subject samples to three freeze-thaw cycles.
- Post-Preparative Stability: Assess stability of processed samples kept in autosampler for 24-72 hours.
5.2 Sample Preparation
- Use at least six replicates for each QC level (LQC and HQC).
- Aliquot QCs from the same batch of plasma to reduce variability.
- Store stability aliquots separately from study samples.
5.3 Execution of Stability Studies
- Label each stability tube with:
- Study code
- Analyte name
- QC level
- Stability type
- Date and time of experiment
- Expose the samples to the defined conditions (e.g., 25°C for 6 hours for short-term).
- Analyze stability samples alongside freshly prepared calibration standards and QCs.
5.4 Data Analysis and Acceptance Criteria
- Calculate measured concentrations of stability samples using fresh calibration curve.
- Acceptance Criteria:
- Mean concentration should be within ±15% of nominal for both LQC and HQC.
- %CV should be ≤15% across replicates.
- Document findings in Annexure-1: Stability Summary Report.
5.5 Documentation and Reporting
- Include raw data printouts, calibration curves, and chromatograms in the stability file.
- Submit final stability report to QA for verification and archival.
- Stability summaries shall be part of the method validation or bioanalytical report.
6. Abbreviations
- BA/BE: Bioavailability/Bioequivalence
- LQC: Low Quality Control
- HQC: High Quality Control
- %CV: Percent Coefficient of Variation
- QC: Quality Control
7. Documents
- Stability Summary Report – Annexure-1
- Raw Data Printouts
- Chromatographic Data Files
8. References
- ICH M10: Bioanalytical Method Validation
- US FDA Guidance for Industry: Bioanalytical Method Validation (2022)
- Sponsor 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: Stability Summary Report
| Type | QC Level | Nominal (ng/mL) | Mean Measured | %CV | % Difference | Status |
|---|---|---|---|---|---|---|
| Short-Term | LQC | 20.0 | 19.2 | 4.8 | -4% | Accepted |
| Freeze-Thaw | HQC | 1500.0 | 1523.4 | 3.2 | +1.6% | Accepted |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 01/01/2022 | 1.0 | Initial issue | New SOP | QA Head |
| 17/04/2025 | 2.0 | Revised layout and included stability types | ICH M10 alignment | QA Head |