Standard Operating Procedure for Bioanalytical Sample Batch Preparation in BA/BE Studies
| Department | BA-BE Studies |
|---|---|
| SOP No. | SOP/BA-BE/168/2025 |
| Supersedes | SOP/BA-BE/168/2022 |
| Page No. | Page 1 of 10 |
| Issue Date | 17/04/2025 |
| Effective Date | 20/04/2025 |
| Review Date | 17/04/2026 |
1. Purpose
To describe the procedure for organizing and preparing batches of plasma samples, calibrators, and quality controls for bioanalytical analysis in BA/BE studies using LC-MS/MS.
2. Scope
This SOP applies to all analysts involved in preparing analytical runs of clinical plasma samples, standards, and QC samples for BA/BE bioanalysis at the bioanalytical laboratory.
3. Responsibilities
- Analyst: Prepares the sample batch layout, labels vials, and compiles
sequence information.
Supervisor: Reviews and approves the batch plan and randomization.
QA: Ensures integrity of randomization and compliance with SOP.
4. Accountability
The Head of Bioanalytical Operations is accountable for ensuring sample batch preparation aligns with study protocol, validated methods, and regulatory standards.
5. Procedure
5.1 Batch Layout and Planning
- Define batch size based on:
- Instrument run time limits (e.g., <36 hrs)
- Total number of subject samples
- Number of replicates and QCs per batch
- Each batch must contain:
- Blank samples (double and single)
- At least 6 non-zero calibrators
- Minimum 3 levels of QC (LQC, MQC, HQC) in duplicate
- Prepare Annexure-1: Batch Preparation Sheet listing injection order and vial positions.
5.2 Randomization and Sample Arrangement
- Use software-generated randomization to assign sample IDs to vial positions.
- Include at least one anchor QC at beginning, middle, and end of the batch.
- Distribute study samples such that:
- No sequential subject samples occur
- Treatment arms are balanced across batches
5.3 Vial Labeling and Loading
- Label all vials (calibrators, QCs, and samples) with printed barcode labels containing:
- Sample ID
- Vial position (e.g., A1, B5)
- Run ID
- Arrange vials in autosampler trays as per layout sheet.
- Record physical arrangement in Annexure-2: Sample Tray Log.
5.4 Sequence File Generation
- Create sequence file using instrument software:
- Define injection order as per batch plan
- Assign compound names, transitions, retention time, and integration parameters
- Save file with study code and batch number.
- Review sequence file and get QA sign-off before starting acquisition.
5.5 Documentation
- All batch preparation documents must be signed and version-controlled.
- Attach randomization file, batch layout, tray map, and sequence file to analytical batch folder.
- Discrepancies must be recorded in Annexure-3: Batch Preparation Deviation Log.
6. Abbreviations
- BA/BE: Bioavailability/Bioequivalence
- LQC/MQC/HQC: Low/Mid/High Quality Control
- QC: Quality Control
- SOP: Standard Operating Procedure
7. Documents
- Batch Preparation Sheet – Annexure-1
- Sample Tray Log – Annexure-2
- Batch Preparation Deviation Log – Annexure-3
8. References
- ICH M10 Guideline for Bioanalytical Method Validation
- US FDA Bioanalytical Method Validation Guidance
9. SOP Version
Version: 2.0
10. Approval Section
| Prepared By | Checked By | Approved By | |
|---|---|---|---|
| Signature | |||
| Date | |||
| Name | |||
| Designation | |||
| Department |
11. Annexures
Annexure-1: Batch Preparation Sheet
| Injection No. | Sample Type | Sample ID | Vial Position |
|---|---|---|---|
| 1 | Blank | BLK | A1 |
| 2 | Calibrator-1 | STD01 | A2 |
| 15 | Subject Sample | BE101-TP3 | C3 |
Annexure-2: Sample Tray Log
| Tray ID | Sample ID | Position | Checked By | Date |
|---|---|---|---|---|
| TR-05 | BE101-TP3 | C3 | Rajesh Kumar | 16/04/2025 |
Annexure-3: Batch Preparation Deviation Log
| Date | Deviation Observed | Corrective Action | Approved By |
|---|---|---|---|
| 16/04/2025 | Wrong vial label for STD03 | Relabeled and re-verified | QA Manager |
Revision History:
| Revision Date | Revision No. | Details | Reason | Approved By |
|---|---|---|---|---|
| 01/01/2022 | 1.0 | Initial release | New SOP | QA Head |
| 17/04/2025 | 2.0 | Inclusion of barcode labeling and deviation log | Compliance update | QA Head |