Procedure for Pyrogen Testing

1) Purpose

The purpose of this SOP is to outline the procedure for performing pyrogen testing on pharmaceutical products to ensure they are free from pyrogenic contaminants and safe for patient use.

2) Scope

This SOP applies to all pharmaceutical products manufactured within the facility that require pyrogen testing as per regulatory requirements.

3) Responsibilities

Quality Control (QC) Department: Responsible for conducting pyrogen testing and ensuring compliance with this SOP.
Microbiology Laboratory: Responsible for performing pyrogen testing procedures and maintaining sterile testing conditions.

4) Procedure

4.1 Equipment and Reagent Preparation:
    4.1.1 Prepare the LAL (Limulus Amebocyte Lysate) reagent as per manufacturer’s instructions.
    4.1.2 Ensure all equipment, including pyrogen-free glassware and endotoxin-free water, is prepared and validated.
    4.1.3 Validate the pyrogen-free status of all materials and reagents used in the testing.

4.2 Sample Collection and Preparation:
    4.2.1 Obtain representative samples of the pharmaceutical product from the production department.
    4.2.2 Transfer samples aseptically into pyrogen-free containers suitable for testing.
    4.2.3 Ensure samples are appropriately labeled and documented to maintain traceability.

4.3 Pyrogen Testing Procedure:
    4.3.1 Perform the LAL test by adding LAL reagent to sample aliquots in pyrogen-free glassware.
    4.3.2 Incubate the samples and LAL reagent mixture under controlled conditions to promote reaction.
    4.3.3 Measure and record the reaction endpoint using appropriate instrumentation or visual assessment.
    4.3.4 Include appropriate positive and negative controls in each test run to validate test performance.

4.4 Calculation and Interpretation:
    4.4.1 Calculate the endotoxin equivalent units (EU) based on the LAL reaction and sample dilutions.
    4.4.2 Compare the measured EU values against acceptance criteria specified in pharmacopeial standards or internal specifications.
    4.4.3 Document and investigate any deviations from expected results, taking corrective actions as necessary.

4.5 Acceptance Criteria:
    4.5.1 Pharmaceutical products are considered pyrogen-free if the EU levels are below specified limits for safe administration.
    4.5.2 Ensure all positive and negative controls demonstrate appropriate responses within established limits.

4.6 Documentation:
    4.6.1 Record all pyrogen testing procedures, results, and observations in the Pyrogen Testing Record.
    4.6.2 Maintain detailed records of sample handling, test conditions, and any deviations encountered.
    4.6.3 Review and approve the documentation by QC Manager.

5) Abbreviations, if any

QC: Quality Control
LAL: Limulus Amebocyte Lysate

6) Documents, if any

Pyrogen Testing Record
Validation Report for Pyrogen Testing Method
Sampling Plan Document

7) Reference, if any

USP General Chapter <85> – Bacterial Endotoxins and Pyrogens Test
Pharmacopeial standards for Pyrogen Testing

8) SOP Version

Version 1.0