The document provides an overview of validation in sterilization processes, emphasizing its importance in ensuring products meet specifications. It outlines principles such as installation, operational, and performance qualifications, and details types of indicators used to verify sterilization effectiveness, including physical, chemical, and biological indicators. Additionally, procedures for testing filter integrity and sterilization efficacy are described, highlighting the significance of biological indicators in confirming sterilization success.

1. Brief Introduction on
Validation
Presented by:
Dr. Malkiet Kaur
MMCP, MM(DU)
Mullana

2. Validation
It is defined as action of checking or proving the validity or accuracy of
something.
OR
It is a established documented evidence which provides high degree of
assurance that specific process will be consistently produce a product
meeting its predetermined specifications and quality attributes.

3. Principles
• Installation qualification: To ensure that equipment used for
sterilization is installed properly.
• Operational qualification: Calibration and testing of instrument is
done to ensure the proper working of the instrument.
• Performance qualification: To ensure the completion of actual
sterilization methods and to check whether the actual conditions are
achieved or not.

4. Indicators
• An indicator is a substance or device used to monitor and verify that the sterilization
process has effectively achieved the desired level of microbial destruction. These
indicators serve as a means of assessing the performance of the sterilization process.
Types of indicators:
• Physical indicators
• Chemical indicators
• Biological indicators

5. Physical indicators
It involves observing the display on the sterilizer and to record the time temperature and pressure.
• DOSIMETER: It is a physical indicator that measures the absorbed dose of ionizing radiation.
For example: Radiation sterilization methods depend on delivering a specific dose of radiation to
achieve sterility. Where, dosimeters confirms that whether the required dose (25kGy for a sterility
assurance level of 10-6
) is delivered to the product load or not.
They are placed at strategic locations within the sterilization load (surface, core and edges) to
measure dose distribution.

6. • Dioctyl Phthalate testing (DOP) testing: It is used for air filtration in LAF.
It is a process in which the integrity of HEPA filter is tested through the
introduction of particulates.
Procedure
• An aerosol (DOP) is introduced upstream of the HEPA filter. The aerosol
particles are usually around 0.3 microns for HEPA filters.
• A photometer measures the aerosol concentration downstream of the filter.
Any particles detected downstream indicate a leak or filter inefficiency.

7. • Bubble pressure test: It is used to validate sterilization filters, such as sterilizing-
grade membrane filters in pharmaceutical and medical device manufacturing.
Purpose:
 To confirm the integrity of sterilization filters.
 To validate that filter can reliably achieve sterility assurance.
 To detect any damage, pinholes or defects in the filter material or sealing.
Calculated by:
D=30γ/P
D= Pore size, γ=Surface tension, p=Pressure

8. Chemical Indicators
These are the substances which gives the visibility by change in color.
1. Process Indicators: It is a simple tool used in sterilization to show whether an item is
exposed to the sterilization process. They are typically placed on the outside of
sterilization packaging like autoclave tape/ sterilization pouches and change in color
when exposed to sterilizing agent (Steam, ethylene oxide or hydrogen peroxide).
2. Bowie-Dick test: Used in steam sterilizers to ensure effective air removal and steam
penetration in the chamber. It contains a chemical indicator sheet inside layers of
porous material. A uniform color change indicates proper air removal, while uneven
results may signal air leaks or vacuum issues.

9. Biological Indicators
• It is a tool used in sterilization processes to verify the effectiveness of the sterilization cycle by assessing its ability
to kill highly resistant microorganisms. They are considered the most reliable method for confirming sterility.
Procedure:
• Bis are placed in sterilizer.
• Sterilization process is run
• Bis are incubated to allow any surviving spores to grow.
• Growth is indicated by a color change in the medium or by turbidity.
• Interpretation: No growth: Indicates successful sterilization.
Growth detected: Indicates the sterilization process was not effective.