BA-BE Studies: SOP for Bioanalytical Sample Batch Preparation – V 2.0

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