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

1. Ensure centrifuged tubes are:
   • Intact without hemolysis or leakage
   • Properly labelled
   • Handled using gloves to avoid contamination
2. Verify each subject ID and time point using the sampling log.

5.2 Plasma Separation

1. Use sterile disposable pipettes for each sample.
2. Carefully aspirate the clear plasma layer without disturbing the buffy coat or red cells.
3. Transfer plasma to pre-labelled cryovials or polypropylene tubes.

5.3 Aliquoting

1. 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
2. Place immediately in cold racks or freezer boxes based on specified storage conditions.

5.4 Sample Volume Documentation

1. Record the volume of plasma separated and aliquoted in Annexure-1: Plasma Separation Log.
2. Note any discrepancies (e.g., low volume, hemolysis) and inform QA and the PI.

5.5 Sample Transfer and Storage

1. Transfer samples to designated storage freezer:
   • -20°C for interim storage
   • -70°C for long-term storage, if specified
2. Log freezer details and time of storage in Annexure-2: Aliquot Storage Log.

5.6 Deviation Handling

1. 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

1. Plasma Separation Log – Annexure-1
2. Aliquot Storage Log – Annexure-2
3. 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