DEPYROGENATION OF GLASSWARE USED FOR BACTERIAL ENDOTOXIN TEST IN MICROBIOLOGY LAB
DEPYROGENATION OF GLASSWARE USED FOR BACTERIAL ENDOTOXIN TEST IN MICROBIOLOGY LAB
Depyrogenation of glassware is a crucial process in pharmaceutical, biotechnology, and other industries where sterile and pyrogen-free glass containers are required. Pyrogens are substances derived from the cell walls of certain microorganisms, such as bacteria, which can cause fever and other adverse reactions when injected into the human body. Therefore, removing pyrogens from glassware is essential to ensure the safety and efficacy of pharmaceutical products, particularly those administered through injections or infusions.
The depyrogenation process involves subjecting glassware, such as vials, ampoules, and syringes, to high temperatures to destroy any pyrogens present. There are two main methods of depyrogenation:
- Dry Heat Depyrogenation: In this method, glassware is exposed to dry heat at temperatures typically ranging from 250°C to 300°C for a specific duration. The high heat effectively breaks down and eliminates pyrogens from the glass surfaces.
- Endotoxin Removal: Endotoxins are a specific type of pyrogens found in the cell walls of Gram-negative bacteria. Depyrogenation methods, such as filtration through specialized membranes or the use of chemical treatments, can effectively remove endotoxins from glassware.
The depyrogenation process must be validated to ensure its effectiveness and compliance with regulatory requirements. Validation involves performing tests to verify that the glassware is free from pyrogens after depyrogenation. These tests may include the Limulus Amebocyte Lysate (LAL) test, which detects the presence of endotoxins, and other specific assays to confirm the absence of pyrogens.
Depyrogenation of glassware is essential to maintain the quality and safety of pharmaceutical products. It ensures that parenteral medications and medical devices are free from pyrogens, reducing the risk of adverse reactions in patients and ensuring the products’ compliance with regulatory standards. By following strict depyrogenation processes and conducting thorough validation, pharmaceutical and biotechnology companies can deliver high-quality, safe, and effective products to patients worldwide.
PURPOSE OF DEPYROGENATION OF GLASSWARE
To provide a procedure for depyrogenation of glassware used for bacterial endotoxin test in Microbiology Lab.
SCOPE
This procedure is applicable for depyrogenation of glassware used for bacterial endotoxin test in Microbiology Lab..
REFERENCE(S) & ATTACHMENTS
REFERENCES
In-house and IP-2022
CROSS REFERENCE DOCUMENTS
Cleaning & Operation of Dry Heat Sterilizer (DHS) in Microbiology Lab.
DEFINITIONS & ABBREVATION(S)
DEFINITIONS
DEPYROGENATION: Depyrogenation is the removal of pyrogen (endotoxins) from the pharmaceutical materials or equipment.
ABBREVIATIONS
QA : Quality assurance
SOP : Standard Operating Procedures
CC No. : Change control number
QC : Quality Control
DHS : Dry Heat Sterilizer
BET : Bacterial Endotoxin Test
RESPONSIBILITY:
Housekeeping Person/Quality control Department person (Microbiologist):
To clean the glass wares for depyrogenation as per respective cleaning procedure.
Microbiologist shall depyrogenate the glass tubes as per procedure.
Head-QC /Designee initiator department shall be responsible for:
To ensure the activity as per SOP & implementation of SOP.
QA Personnel shall be responsible for:
To review the SOP and distribute the control copy to department.
Head-QA/Designee shall be responsible for:
To ensure the implementation & compliance of the defined procedure.
Plant Head/ Designee shall be responsible for:
To ensure the implementation of the procedure.
PROCEDURE:
SAFETY/PRECAUTION/EHS
Always wear thermo-resistant gloves to unload the glassware after depyrogenation process.
Temperature of the DHS should be regularly monitored at certain intervals during the Depyrogenation process, if it is found out of the limit, do not use the glassware of this lot for BET.
Unwrap the depyrogenated glassware only before the performing the BET. If any depyrogenated item is found without foil or foil is damaged, discard such glassware and don’t use the same for BET.
Procedure
All the glassware including BET tubes and dilution tubes for BET shall be cleaned..
After completion of drying, remove the glassware from the drying oven and let it attain the room temperature.
Properly wrap the glassware with aluminum foil for depyrogenation.
Put all these wrapped glassware on the perforated shelves of dry heat sterilizer (DHS) with keeping sufficient space between each wrapped bundle as per validated cycle of glassware for the proper heat distribution.
Operate the Dry Heat Sterilizer at 250±50C for 30 minutes.
After completion of depyrogenation process open the door of DHS and placed the glassware on the respective place with help of heat resistant gloves.
Use these glassware for BET testing.