Standard Operating Procedure for Short-Term Stability of Analyte in BA/BE Studies
| Department | BA-BE Studies |
|---|---|
| SOP No. | SOP/BA-BE/175/2025 |
| Supersedes | SOP/BA-BE/175/2022 |
| Page No. | Page 1 of 9 |
| Issue Date | 17/04/2025 |
| Effective Date | 20/04/2025 |
| Review Date | 17/04/2026 |
1. Purpose
To define the procedure for conducting short-term stability studies of analytes in plasma to ensure sample integrity during temporary exposure to room temperature conditions during sample processing in bioavailability/bioequivalence (BA/BE) studies.
2. Scope
This SOP applies to the assessment of short-term stability of plasma samples stored temporarily at room temperature (~25°C) during processing and prior to injection into the LC-MS/MS system in BA/BE studies.
3. Responsibilities
- Analyst: Prepares and processes QC samples for short-term stability study.
- Reviewer: Verifies the results and compliance with acceptance criteria.
- QA Officer: Reviews and approves documentation and ensures adherence to regulatory guidance.
4. Accountability
The Head of the Bioanalytical Department is accountable for ensuring correct execution, interpretation, and documentation of short-term stability data in accordance with ICH M10 and other applicable guidelines.
5. Procedure
5.1 Sample Preparation
- Prepare a minimum of six replicates of Low QC (LQC) and High QC (HQC) levels.
- Aliquot the prepared samples in appropriately labeled tubes indicating:
- Study Code
- Analyte
- QC Level
- Short-Term Stability
- Date and Time
- Store one set under standard frozen condition (control) and another set at room temperature (~25°C).
5.2 Exposure Duration
- Keep stability sample set at ambient temperature for a pre-defined duration:
- Typically 4, 6, or 24 hours depending on method validation protocol.
- After the exposure period, return the samples to -20°C until analysis.
5.3 Sample Analysis
- Analyze short-term exposed samples with freshly prepared calibration standards and frozen control QCs using validated LC-MS/MS method.
- Document retention time, area counts, and concentration of each replicate.
5.4 Acceptance Criteria
- Mean measured concentration of LQC and HQC should be within ±15% of nominal values.
- %CV should be ≤15% across replicates at each QC level.
- Record and compare results in Annexure-1: Short-Term Stability Summary.
5.5 Reporting
- Document observations and outcomes in the short-term stability report.
- Submit report to QA for review and approval.
- Include stability report as part of the method validation or study report.
6. Abbreviations
- BA/BE: Bioavailability/Bioequivalence
- LQC: Low Quality Control
- HQC: High Quality Control
- %CV: Percent Coefficient of Variation
- QA: Quality Assurance
7. Documents
- Short-Term Stability Summary – Annexure-1
- Chromatographic Data and Calibration Curves
8. References
- ICH M10: Bioanalytical Method Validation
- US FDA Guidance for Bioanalytical Method Validation
- 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: Short-Term Stability Summary
| QC Level | Nominal (ng/mL) | Mean Measured | %CV | % Difference | Status |
|---|---|---|---|---|---|
| LQC | 20.0 | 19.1 | 5.2 | -4.5% | Accepted |
| HQC | 1500.0 | 1492.3 | 2.7 | -0.5% | Accepted |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 01/01/2022 | 1.0 | Initial SOP Issue | New Procedure | QA Head |
| 17/04/2025 | 2.0 | Updated to align with ICH M10 and clarified %CV requirement | Regulatory Update | QA Head |