reproducible chromatographic results.

2. Scope

This SOP applies to all chromatographic columns (HPLC, UHPLC, GC) used in analytical method development laboratories for qualitative and quantitative analysis of pharmaceutical substances and drug products.

3. Responsibilities

• Analytical Chemist: Responsible for performing the qualification and documenting the results.
• Section Supervisor: Verifies qualification protocol adherence and checks calculations.
• QA Officer: Reviews and approves the qualification report and ensures compliance with GMP requirements.

4. Accountability

The Head of Analytical Method Development is accountable for the implementation, review, and periodic update of this SOP and ensuring all columns are qualified prior to use.

5. Procedure

5.1 Column Receipt and Entry

1. Receive the column from an approved vendor along with Certificate of Analysis (CoA).
2. Log the column into the Column Inventory Register (Annexure-1) with details including:
   • Column type and dimensions
   • Serial number and batch number
   • Manufacturer and part number
   • Date of receipt and user name

5.2 Qualification Criteria

1. Use the system suitability method of an existing validated analytical procedure that employs the same column type.
2. Perform injections of standard solution in replicates (minimum 5 injections).
3. Evaluate the following parameters:
   • Retention time (%RSD ≤ 1.0%)
   • Theoretical plates (N) ≥ minimum specified in method
   • Tailing factor ≤ 2.0
   • Resolution ≥ 2.0 between critical pairs (if applicable)
4. Flush column with manufacturer-recommended solvents prior to and after testing.

5.3 Data Compilation and Evaluation

1. Record chromatograms and instrument reports.
2. Fill in the Chromatographic Column Qualification Report (Annexure-2).
3. Compare the results with predefined acceptance criteria.
4. Investigate any non-compliance and perform root cause analysis if necessary.

5.4 Column Tagging and Release

1. Tag qualified columns with a unique identifier and qualification status (Qualified / Rejected).
2. Update the Column Inventory Register with status and qualification date.
3. Store qualified columns in designated column cabinet under controlled conditions.

5.5 Requalification

1. Requalification shall be done:
   • After 100 injections
   • If peak shape anomalies or resolution issues are observed
   • Following long-term storage (>6 months)
2. Follow the same procedure as initial qualification.

6. Abbreviations

• HPLC: High-Performance Liquid Chromatography
• GC: Gas Chromatography
• RSD: Relative Standard Deviation
• QA: Quality Assurance
• CoA: Certificate of Analysis

7. Documents

1. Column Inventory Register – Annexure-1
2. Chromatographic Column Qualification Report – Annexure-2

8. References

• ICH Q2(R1) – Validation of Analytical Procedures
• USP <621> – Chromatography
• 21 CFR Part 211 – Current Good Manufacturing Practice

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Ankita Sharma	Rahul Menon	Dr. Preeti Verma
Designation	Analyst	QA Officer	Head – AMD
Department	Analytical Method Development	Quality Assurance	Analytical Method Development

11. Annexures

Annexure-1: Column Inventory Register

Sr. No.	Column Type	Serial No.	Date of Receipt	Status
001	C18, 250×4.6mm, 5μ	12345678	25/05/2025	Qualified

Annexure-2: Chromatographic Column Qualification Report

Includes injection sequence, retention time, tailing factor, theoretical plates, resolution data, chromatograms, and remarks.

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/06/2025	2.0	Added requalification conditions and clarified criteria	Periodic Review	Dr. Preeti Verma
12/07/2022	1.0	Initial issue	New SOP	QA Head