AUTOCLAVE

TECHNICAL NOTE

Steam Sterilization Validation 4

Sanjeev patel
sanjeev@gampservices.com
A Brief Overview of Steam Sterilization Cycles:
Cycle development is the process of determining the physical properties
to be met in a sterilization cycle that will be used to reproducibly and
consistently sterilize the product, component, and/or equipment in a
defined loading pattern.

The goal of cycle development is to provide a proven acceptable range

of sterilizing conditions that result in a product/material that is both
sterile and functional after the completion of the sterilization cycle.

Cycle development is an important part of both the autoclave

procurement and validation processes.

Cycle development is performed to provide you with a sterilization

process and recipe that works for your load. This process will result in
parameters (i.e. cycle type, temperature, time, etc.) that can be validated.

Ideally, cycle development should begin before the sterilizer is

purchased, since the cycle types will affect the type of autoclave you
need, and changes in the field after installation are generally difficult
and expensive.

At the very least, cycles should be developed prior to performing a full

validation of the autoclave and, more specifically, before Performance
Qualification (PQ) validation.
Process:
1. Determine cycle type: gravity, prevac, liquids, etc.

2. Once the cycle type is determined you must decide on the key cycle
parameters: sterilization temperature and sterilizer time.
3. Once you have determined your cycle type and parameters, you
must validate the cycle to ensure the load will be sterile.

4. Sterility testing is confirmed with the use of biological indicators

(BIs). The most common test organism for steam sterilization is
Geobacillus stearothermophilus. BIs are placed in intimate contact
with the load and are placed throughout the chamber. For dry loads,
spore strips are used. For liquid loads, you either use a hermetically-
sealed ampoule BI suspended in the liquid with a string or a wire, or
you can inoculate the actual liquid load with a challenge organism
 (Geobacillus stearothermophilus) and culture it after the cycle is
complete.

5. Typically, at least 10 BIs are used per cycle. If all BIs show no growth
(i.e. all the bacteria are destroyed) in three successive cycles, you
have validated the cycle.

All loads cannot be run with the same sterilization cycle. This is why
autoclaves are equipped to run various cycle types.

The following is a list of the most typical steam sterilization cycles

used in laboratories, healthcare, and pharmaceutical production.
Gravity and Liquids
Gravity and Liquids cycles both preheat the load (through gravity
displacement), raise the temperature by admitting steam to the
chamber, and hold the temperature constant (more or less)
throughout the exposure phase of the cycle. The difference
between the two is in what happens after the Sterilization phase.
Gravity cycles vent steam rapidly to the drain and may evacuate the
chamber to dry the load. Liquids cycles slowly drop the pressure
and temperature to ensure that the liquid load does not boil over as
it cools. Liquids cycles may use a load probe to measure and
control the Sterilization phase of the cycle from within the actual
liquid load.
Vacuum
This cycle type is similar to a Gravity cycle, except for what
happens before the Sterilization phase. Multiple pulses of vacuum
followed by steam injection are used to remove air from the
chamber and the load. This is the cycle type of choice for
cannulated instruments, tubing, plumbing, animal bedding, and
anything complex that presents a challenge to steam getting to the
surfaces to be sterilized. It is never to be used with liquid loads.
Air-Over-Pressure

This cycle type is similar to a Liquids cycle except that it injects

compressed air into the chamber at the Sterilization phase to
maintain pressure during cool down. This cycle type is best used
for small-volumes of liquid loads that tend to boil over.
Developing a Sterilization Cycle:
Step 1: Determine the Cycle Type
Which cycle do you use? Refer to the flow-chart below to help you
determine which cycle type is best for your load.

Step 2: Determine the Cycle Parameters: Time and Temperature

Once the cycle type is determined you must decide on the key
cycle parameters: sterilization temperature and sterilization time.
The vast majority of loads sterilized in the laboratory setting are
sterilized at 121.1°C/250°F for 30-90 minutes.

If your load contains items that are sensitive to heat, the default
cycle temperature of 121.1°C/250°F may be too hot, and you might
need to consider a lower temperature. Using lower sterilization
temperatures will result in longer sterilization times. This can be
calculated by using the F0 relationship:
F0 is the accumulated lethality in minutes at 121.1°C/250°F.
t = the amount of time in minutes of the sterilization phase of the sterilization
cycle.
T = the sterilization temperature in °C.
The F0 equation solves to:

Let’s take an example by running the numbers for a sterilization cycle using
110°C as the sterilization temperature. To solve for t (time), plug in an F0 value
of 15 along with the desired T (110°C) on the right side of the equation:

t = 193 minutes
As you can see, lower temperature cycles take more time to achieve
sterilization. As a comparison, at 121.1°C, this cycle would theoretically take
15 minutes to achieve sterilization.

Step 3: Validate the Cycle

Once you have taken a “best guess” at your cycle type, sterilization
temperature, and sterilization time, you must validate the cycle to ensure the
load will be sterile. These test cycles should be loaded in a way that is
representative of the actual load to be sterilized. It is recommended to run your
test cycles at half of the desired sterilization time (this is referred to as the
“half-cycle” method).