Standard Operating Procedure for Chromatogram Review and Analysis in Biosimilar Manufacturing

Department	Biosimilars
SOP No.	SOP/BS/189/2025
Supersedes	SOP/BS/189/2022
Page No.	Page 1 of 9
Issue Date	04/05/2025
Effective Date	06/05/2025
Review Date	04/05/2026

1. Purpose

To establish a standard procedure for reviewing and analyzing chromatograms obtained from biosimilar purification processes and quality control testing, ensuring accurate interpretation and GMP-compliant data reporting.

2. Scope

This SOP applies to all chromatographic data generated from High-Performance Liquid Chromatography (HPLC), Ultra-High Performance Liquid Chromatography (UHPLC), and other related techniques used in biosimilar manufacturing and QC operations.

3. Responsibilities

• QC Analyst: Review chromatograms, integrate peaks, and assess system suitability per method validation criteria.
• QA Reviewer: Cross-check reviewed chromatograms for compliance with specifications and approve or reject based on results.
• Instrumentation Specialist: Maintain chromatographic systems and verify calibration status before use.

4. Accountability

The Quality Control Manager is accountable for ensuring that chromatographic data are reviewed in a timely, accurate, and traceable manner in compliance with GMP and data integrity guidelines.

5. Procedure

5.1 Pre-Review Checklist

1. Ensure chromatographic system is qualified and in calibrated state.
2. Confirm analytical method number, SOP reference, column type, and mobile phase batch number.
3. Verify analyst, date, and sample ID recorded in the raw data file.

5.2 System Suitability Check

1. Review system suitability chromatogram for parameters such as:
   • Resolution between peaks
   • Tailing factor (should be < 2.0)
   • Theoretical plates (N)
   • Retention time consistency
2. If suitability fails, do not proceed with sample analysis. Investigate per deviation SOP.

5.3 Chromatogram Evaluation

1. Open raw data in validated chromatography software (e.g., Empower, Chromeleon).
2. Verify baseline placement and correct peak integration.
3. Ensure consistent integration settings across samples and standards.
4. Assign peak identities using retention time and method-defined criteria.

5.4 Quantitative Analysis

1. Confirm correct use of calibration curve and response factor.
2. Ensure sample dilution, injection volume, and integration method match the BMR or analytical method.
3. Calculate analyte concentrations and report with units (e.g., mg/mL, % purity).

5.5 Documentation and Approval

1. Print chromatograms with audit trail and relevant overlays (standard/sample).
2. Sign and date chromatogram, attach review comments.
3. Submit for QA review and archive according to data integrity policy.

6. Abbreviations

• HPLC: High-Performance Liquid Chromatography
• QC: Quality Control
• RT: Retention Time
• SST: System Suitability Test

7. Documents

1. Chromatogram Review Log – Annexure-1
2. System Suitability Checklist – Annexure-2
3. Integration and Calculation Record – Annexure-3

8. References

• ICH Q2(R2) – Validation of Analytical Procedures
• FDA Guidance – Data Integrity and Compliance with cGMP
• WHO TRS 1004 – GMP for Biotherapeutics

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Chromatogram Review Log

Date	Sample ID	Reviewer	Software	Comments
04/05/2025	BS-CHR-011	Ajay Verma	Empower	Peak tailing acceptable

Annexure-2: System Suitability Checklist

Parameter	Acceptance Criteria	Observed	Status
Resolution	> 2.0	2.4	Pass
Tailing Factor	< 2.0	1.5	Pass
Retention Time	±2%	1.98%	Pass

Annexure-3: Integration and Calculation Record

Sample ID	Peak Area	Response Factor	Concentration	Reviewed By
BS-SM-121	843256	1.12	9.8 mg/mL	Sunita Reddy

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
04/05/2025	2.0	Updated system suitability limits and chromatographic software references	Compliance Alignment