Standard Operating Procedure (SOP)

1. Purpose

To establish a standardized procedure for performing System Suitability Testing (SST) in High-Performance Liquid Chromatography (HPLC) analysis to ensure system integrity, performance, and reliable analytical results.

2. Scope

This SOP applies to all HPLC systems and related analyses conducted in [insert laboratory/department/organization name].

3. Responsibility

• Analysts: Responsible for performing SST prior to analysis.

• QC Supervisor/Lab Manager: Responsible for reviewing SST data and ensuring compliance.

• QA Department: Responsible for periodic audits and SOP compliance.

4. Definitions

• System Suitability Test (SST): A set of tests to verify that the HPLC system is working properly before or during sample analysis.

• HPLC: High-Performance Liquid Chromatography, a technique used to separate, identify, and quantify components in a mixture.

5. Materials and Equipment

• HPLC System (Pump, Autosampler, Detector, Column, Software)

• SST Standard (e.g., USP reference standard, or in-house validated standard)

• Mobile Phase

• Column (as per method)

• Data acquisition system (chromatography software)

6. System Suitability Parameters (Typical Criteria)

Note: Use method-specific criteria if specified in pharmacopeial monographs or validated methods.

7. Procedure

7.1 Preparation

1. Ensure that the HPLC system is powered on, equilibrated, and properly flushed with the mobile phase.

2. Install the correct column and allow it to equilibrate with the mobile phase for at least 30 minutes (or as per method).

3. Prepare the SST solution as per validated method or pharmacopeial requirements.

7.2 Running the SST

1. Inject a minimum of 5 replicates of the SST standard into the system.

2. Record chromatographic parameters such as retention time, area, tailing factor, resolution, and theoretical plates.

7.3 Evaluation of SST Results

1. Retention Time: Check for consistency between injections (within ±2%).

2. %RSD of Peak Area: Calculate the %RSD from 5 replicate injections. Should not exceed 2.0% (unless otherwise specified).

3. Tailing Factor: Evaluate for the main peak; must be ≤2.0.

4. Resolution: Ensure Rs ≥2.0 between closely eluting peaks.

5. Theoretical Plates: Calculate efficiency (N) for main peak; it should be within method-specific limits.

7.4 Documentation

• Record all SST data in the chromatography software and maintain printed or electronic records.

• Complete the system suitability checklist or log sheet (if applicable).

• If SST fails, investigate, troubleshoot, and re-perform the test.

8. Acceptance Criteria and Action

• If SST meets criteria: Proceed with sample analysis.

• If SST fails:

  • Do not analyze samples.

  • Investigate possible causes (e.g., mobile phase, column, detector, system pressure).

  • Document the issue in the lab incident log.

  • Correct the issue, re-run SST, and only proceed if it passes.

9. Troubleshooting Guidelines

10. Records and Documentation

• Maintain all SST data with chromatograms.

• SST logbook or electronic records should include:

  • Date

  • Method Name

  • Analyst Name

  • HPLC System ID

  • Standard used

  • SST parameters and results

  • Any deviations and corrective actions

11. Frequency

• SST must be performed before every analytical run.

• Also required after system maintenance, column change, or method change.

12. References

• USP <621> Chromatography

• ICH Q2(R1) – Validation of Analytical Procedures

• Instrument Manufacturer’s Manual.

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