Standard Operating Procedure for Plasma Separation and Aliquoting in BA/BE Studies
| Department | BA-BE Studies |
|---|---|
| SOP No. | SOP/BA-BE/074/2025 |
| Supersedes | SOP/BA-BE/074/2022 |
| Page No. | Page 1 of 10 |
| Issue Date | 17/04/2025 |
| Effective Date | 20/04/2025 |
| Review Date | 17/04/2026 |
1. Purpose
To define the procedure for separation of plasma from whole blood and its subsequent aliquoting into labelled containers for storage and bioanalysis during Bioavailability/Bioequivalence (BA/BE) studies.
2. Scope
This SOP applies to all laboratory personnel involved in the post-centrifugation handling of blood samples for plasma separation and aliquoting at the clinical research facility or designated laboratory site.
3. Responsibilities
- Lab Technician: Responsible for performing plasma separation and aliquoting as per protocol instructions.
- Clinical Research Coordinator (CRC): Ensures sample labels are verified and matched with subject schedule and timepoint.
- QA Personnel: Reviews log entries and verifies sample integrity and compliance during audits.
4. Accountability
The Bioanalytical Manager is accountable for ensuring plasma separation and aliquoting are conducted in accordance with regulatory requirements and analytical reliability standards.
5. Procedure
5.1 Pre-Separation Checks
- Ensure centrifuged tubes are:
- Intact without hemolysis or leakage
- Properly labelled
- Handled using gloves to avoid contamination
- Verify each subject ID and time point using the sampling log.
5.2 Plasma Separation
- Use sterile disposable pipettes for each sample.
- Carefully aspirate the clear plasma layer without disturbing the buffy coat or red cells.
- Transfer plasma to pre-labelled cryovials or polypropylene tubes.
5.3 Aliquoting
- If multiple aliquots are required:
- Equally divide the plasma into pre-labelled aliquot tubes (e.g., A1, A2, A3)
- Label tubes with subject ID, timepoint, aliquot number, and date
- Place immediately in cold racks or freezer boxes based on specified storage conditions.
5.4 Sample Volume Documentation
- Record the volume of plasma separated and aliquoted in Annexure-1: Plasma Separation Log.
- Note any discrepancies (e.g., low volume, hemolysis) and inform QA and the PI.
5.5 Sample Transfer and Storage
- Transfer samples to designated storage freezer:
- -20°C for interim storage
- -70°C for long-term storage, if specified
- Log freezer details and time of storage in Annexure-2: Aliquot Storage Log.
5.6 Deviation Handling
- Any cross-contamination, incorrect labelling, or sample damage must be documented in Annexure-3: Sample Handling Deviation Log.
6. Abbreviations
- BA: Bioavailability
- BE: Bioequivalence
- CRC: Clinical Research Coordinator
- QA: Quality Assurance
- ID: Identification
7. Documents
- Plasma Separation Log – Annexure-1
- Aliquot Storage Log – Annexure-2
- Sample Handling Deviation Log – Annexure-3
8. References
- ICH E6(R2) – Good Clinical Practice
- FDA BA/BE Guidance for Industry
- Study-Specific Protocol and Lab Manual
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Plasma Separation Log
| Subject ID | Timepoint | Volume (mL) | Aliquot Nos. | Initials | Remarks |
|---|---|---|---|---|---|
| VOL-074 | 2h | 4.5 | A1, A2 | Sunita Reddy | — |
Annexure-2: Aliquot Storage Log
| Subject ID | Aliquot No. | Freezer ID | Rack No. | Storage Temp | Time | Handled By |
|---|---|---|---|---|---|---|
| VOL-074 | A1 | FZ-03 | R2 | -70°C | 09:15 | R. Pawar |
Annexure-3: Sample Handling Deviation Log
| Subject ID | Issue | Date | Reported By | Corrective Action | Reviewed By |
|---|---|---|---|---|---|
| VOL-074X | Hemolysis observed | 17/04/2025 | Nisha Verma | Sample discarded, noted in CRF | Dr. Arvind Shah |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 12/01/2022 | 1.0 | Initial release | Process standardization | QA Head |
| 17/04/2025 | 2.0 | Incorporated detailed volume logging and deviation annexure | Regulatory alignment | QA Head |