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Mastering Sterilization: The Science and Technology Behind Autoclaves

Aishwarya Nair, Biomedical Engineer
7 minutes Read. 16th Jul 2024

Autoclaves play a crucial role in effectively autoclave sterilization eradicating bacteria, viruses, fungi, and spores present on instruments and items. They operate by using pressurized steam, which, when applied over a specific duration, alters, and coagulates microorganisms, rendering them nonviable.

Steam sterilization using autoclaves is a fundamental safety practice in numerous laboratories and medical facilities. Sterilization is essential for removing pathogens from equipment to prevent their transmission. Autoclaves are commonly employed in medical and dental labs to sterilize instruments, ensuring aseptic conditions. Moreover, in the disposal of medical waste, autoclaving is a common step before discarding it. Surgical equipment is also routinely subjected to autoclave sterilization, particularly in healthcare settings. Additionally, autoclaves find application in research and scientific environments for the sterilization of culture media, sample tubes, and pipette tips.

Types of Autoclave sterilization

The need for effective equipment autoclave sterilization drives the choice of methods. Three main sterilization methods are prevalent: steam, dry heat, and ethylene oxide (EtO). Understanding how each method works, along with its pros and cons, is crucial for making informed decisions on the most suitable sterilization approach.

Steam sterilization:

This is also known as autoclaving, is a fast, safe, and cost-effective method for sterilization. It uses high temperatures and pressure to destroy microorganisms and bacterial spores, requiring the maintenance of steam at around 120 degrees Celsius for a minimum of thirty minutes.

Dry heat sterilization

This is a thermal autoclave sterilization processing method that removes moisture content from materials. It is recommended for materials that may be damaged by moist heat or are impermeable to it, such as powders or sharp instruments.

Ethylene Oxide (EtO) Sterilization:

ETO autoclave sterilization is a common method that uses gas to disrupt cell growth and kill microorganisms. It offers advantages such as suitability for heat or moisture-sensitive items, ability to sterilize various materials, and packaging penetration; it comes with significant safety concerns.

How Does an Autoclave Work?

Autoclaves function similarly to pressure cookers. They seal, remove air, and replace it with steam, then pressurize to sterilize. The process includes:

The fundamental principle of steam sterilization (autoclave) involves placing a clean medical device inside an enclosed environment filled with steam, ensuring specific conditions of heat, temperature, pressure, and duration. Autoclaves rely on these four essential criteria to achieve effective sterilization.

  • Purge Phase: The steam displaces air as temperature and pressure rise slightly.
  • Exposure (Sterilization) Phase: Exhaust valve closes, raising temperature and pressure to the setpoint, maintaining it until sterilization is complete.
  • Exhaust Phase: Pressure is released through an exhaust valve, restoring ambient pressure while contents remain hot.

High-quality steam is essential for effective autoclave sterilization, with a recommended composition of 97% steam and 3% moisture for efficient heat transfer. Steam with less than 3% moisture is considered superheated and is not suitable for sterilization due to its inadequate heat transfer capabilities.

5 autoclave sterilization steps

Safety Gear: Prioritize safety with appropriate protective gear for handling infectious materials during biomedical waste disposal and equipment sterilization.

Sterilization Temperature: Autoclaves use steam at 121°C to 134°C for effective sterilization.

Preparation: Before starting, check the autoclave for drain screen blockages and ensure the reservoir is filled with deionized water as needed.

Equipment Inspection: Inspect medical equipment thoroughly before placing it in the autoclave to prevent damage.

Door Safety: Only open the autoclave door after the internal pressure has reached zero and the temperature has cooled below 121°C to avoid steam-related injuries.

Parameter Verification: Verify that the autoclave meets sterilization parameters for safety and effectiveness.

What are the types of autoclaves?

  • Pressure Cooker Type (N-type): They are suitable for small set ups and smaller loads.
  • Gravity Displacement Type: They are commonly used in research laboratories; it displaces air through a drain port using steam.
  • Positive Pressure Displacement Type (B-type): This type is an advanced autoclave that generates steam separately and passes it into the chamber for sterilization.
  • Negative Pressure Displacement Type (S-type): The most expensive type with a vacuum generator and steam generator for thorough sterilization.

What are the benefits of Autoclave Sterilization?

The steam sterilization process offers several benefits, including non-toxicity and ease of control. It relies on key parameters such as drying, air removal, steam contact, time, moisture, temperature, and pressure. Achieving accurate measurements, especially for temperature and pressure, is essential for successful sterilization according to standards like EN285.

Air Removal: 

To ensure saturated steam conditions, air must be effectively removed from the sterilizer chamber and load using a vacuum system, minimizing the presence of non-condensable gases.

Temperature and pressure: 

They are closely linked and can be used to verify the performance of an autoclave. Pressure can be converted into theoretical temperature, allowing a comparison with the actual temperature. This assessment helps ensure that the steam is saturated.

Time: 

Bacteria require a minimum time to be eradicated effectively. The concept of lethality value illustrates the correlation between time and temperature in achieving the same killing effect. Adjusting the exposure time and temperature allows for flexibility in achieving the desired sterilization effect. Choosing a higher temperature, like 134°C, can significantly reduce the required time for autoclave sterilization if the product can withstand higher temperatures. The sterility assurance level (SAL) is a critical consideration, with typical definitions of sterility around 1/1,000,000, ensuring that only one in a million bacteria would survive the sterilization process.

Steam contact: 

This is vital in ensuring that the stored energy in steam is effectively transferred to the contaminated surface for sterilization.

Drying: 

Proper and thorough drying is essential to prevent re-contamination of the sterilized load, typically achieved by applying a vacuum at the end of the cycle to remove condensates.

Moisture: 

The presence of adequate moisture in clean steam is crucial for effective protein denaturation (coagulation). Avoiding superheated steam, which lacks sufficient moisture for proper sterilization, is essential to the process's success.

Autoclaves are silent guardians of safety and hygiene, neutralizing harmful microorganisms through steam, pressure, and time. Their crucial role in sterilization processes spans across industries, from hospitals to research labs, ensuring our well-being by eliminating invisible threats.

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