criteria.

QA Officer: Reviews column qualification records and investigates deviations or failures.

Lab Manager: Ensures traceability of column usage and qualification records.

4. Accountability

The Head of Bioanalytical Department is accountable for ensuring proper qualification, performance monitoring, and replacement of columns in accordance with this SOP.

5. Procedure

5.1 Column Receipt and Inventory

1. Receive column from approved vendor with Certificate of Analysis (CoA).
2. Verify physical condition and label with a unique Column ID.
3. Record details in Annexure-1: Column Inventory Log including:
   • Column type
   • Serial number
   • Dimensions
   • Lot number
   • Date of receipt

5.2 Initial Column Qualification

1. Flush the column with recommended solvents (e.g., methanol, water, acetonitrile) to remove shipping preservatives.
2. Equilibrate the column with mobile phase used in the validated method.
3. Run 5 replicates of a standard solution or system suitability solution.
4. Evaluate performance parameters:
   • Retention time consistency (RSD ≤ 2%)
   • Theoretical plates ≥ method-specified limit
   • Tailing factor ≤ 2.0
   • Resolution between analyte and IS ≥ 2.0
5. Document results in Annexure-2: Column Qualification Report.

5.3 Routine Column Usage and Monitoring

1. Log every use of the column including:
   • Date of use
   • Run ID
   • Number of injections
   • Analyst name
2. Monitor performance weekly or every 500 injections (whichever comes first).
3. Check chromatograms for:
   • Peak symmetry
   • Baseline noise
   • Drift or pressure changes
4. Document observations in Annexure-3: Column Usage and Monitoring Log.

5.4 Column Regeneration and Disposal

1. Regenerate columns by back-flushing with strong solvent if performance declines.
2. If regeneration fails or column is damaged:
   • Mark column as ‘Out of Use’
   • Dispose as per lab’s hazardous waste SOP
   • Record in Annexure-4: Column Disposition Log

6. Abbreviations

• BA/BE: Bioavailability/Bioequivalence
• LC-MS/MS: Liquid Chromatography Tandem Mass Spectrometry
• IS: Internal Standard
• RSD: Relative Standard Deviation
• SOP: Standard Operating Procedure

7. Documents

1. Column Inventory Log – Annexure-1
2. Column Qualification Report – Annexure-2
3. Column Usage and Monitoring Log – Annexure-3
4. Column Disposition Log – Annexure-4

8. References

• ICH M10: Bioanalytical Method Validation
• US FDA Bioanalytical Method Validation Guidance (2022)
• USP General Chapter <621>: Chromatography

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Column Inventory Log

Column ID	Type	Dimensions	Lot No.	Received Date
COL-BAE-001	C18	50×2.1mm, 3µm	LT202504	12/04/2025

Annexure-2: Column Qualification Report

Column ID	RT RSD (%)	Plates	Tailing Factor	Resolution	Status
COL-BAE-001	1.2%	6500	1.1	2.5	Qualified

Annexure-3: Column Usage and Monitoring Log

Date	Run ID	No. of Injections	Observations	Initials
15/04/2025	BAE-102	96	Normal performance	RK

Annexure-4: Column Disposition Log

Column ID	Disposed Date	Reason	Disposed By	Verified By
COL-BAE-001	15/10/2025	High backpressure	Sunita Reddy	QA Officer

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/01/2022	1.0	Initial Issue	New SOP	QA Head
17/04/2025	2.0	Enhanced qualification and lifecycle tracking	Regulatory Update	QA Head