Standard Operating Procedure for Auto-sampler Stability of Analyte in BA/BE Studies
| Department | BA-BE Studies |
|---|---|
| SOP No. | SOP/BA-BE/178/2025 |
| Supersedes | SOP/BA-BE/178/2022 |
| 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 process for evaluating the auto-sampler stability of analytes in processed plasma samples stored in the instrument tray during LC-MS/MS runs in bioavailability/bioequivalence (BA/BE) studies.
2. Scope
This SOP is applicable to all validated bioanalytical methods used in BA/BE studies, specifically for assessing the stability of analytes held at auto-sampler temperature (usually 4–10°C) for prolonged durations before injection.
3. Responsibilities
- Bioanalytical Analyst: Prepares and analyzes auto-sampler stability samples and maintains logs.
- Supervisor: Ensures compliance with time schedules and evaluates results.
- QA Officer: Reviews documentation and approves inclusion in validation or study report.
4. Accountability
The Bioanalytical Head is accountable for ensuring the auto-sampler stability studies are conducted according to validation protocols and documented accurately for regulatory compliance.
5. Procedure
5.1 Sample Preparation
- Prepare a minimum of six replicates each for Low QC (LQC) and High QC (HQC) samples.
- Label each vial clearly with:
- Sample ID
- QC Level
- Analyte Name
- Preparation Date & Time
- Auto-sampler Stability
- Record sample details in Annexure-1: Auto-sampler Stability Log.
5.2 Exposure in Auto-sampler
- Place processed QC samples in the auto-sampler tray set to validated temperature (e.g., 4°C).
- Expose samples for a predefined duration (e.g., 24, 48, or 72 hours) as specified in the method validation protocol.
- Ensure samples remain capped and undisturbed throughout the holding period.
5.3 Sample Analysis
- Inject stability samples along with fresh calibration curve and system suitability QC samples.
- Use validated LC-MS/MS method and record analyte concentrations.
5.4 Acceptance Criteria
- Mean concentrations of LQC and HQC must be within ±15% of their nominal values.
- %CV for each QC level must be ≤15%.
- Summarize findings in Annexure-2: Auto-sampler Stability Summary.
5.5 Documentation and Reporting
- Compile chromatograms, raw data printouts, and analysis reports.
- Prepare and submit a report to QA for approval.
- Include the approved report in the study validation or final submission dossier.
6. Abbreviations
- BA/BE: Bioavailability/Bioequivalence
- LQC: Low Quality Control
- HQC: High Quality Control
- %CV: Percent Coefficient of Variation
- QA: Quality Assurance
7. Documents
- Auto-sampler Stability Log – Annexure-1
- Auto-sampler Stability Summary – Annexure-2
8. References
- ICH M10: Bioanalytical Method Validation
- US FDA Guidance for Industry: Bioanalytical Method Validation (2022)
- Internal 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: Auto-sampler Stability Log
| Sample ID | QC Level | Prep Date | Tray Temp | Start Time | End Time | Analyst |
|---|---|---|---|---|---|---|
| BE178-AS-01 | LQC | 16/04/2025 | 4°C | 10:00 | 10:00 (18/04/2025) | Rajesh Kumar |
Annexure-2: Auto-sampler Stability Summary
| QC Level | Nominal (ng/mL) | Mean Measured | %CV | % Difference | Status |
|---|---|---|---|---|---|
| LQC | 20.0 | 19.2 | 4.3 | -4.0% | Accepted |
| HQC | 1500.0 | 1496.5 | 3.1 | -0.2% | Accepted |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 01/01/2022 | 1.0 | Initial SOP Issuance | New SOP | QA Head |
| 17/04/2025 | 2.0 | Updated cycle durations, annexures added | Method Compliance | QA Head |