GUI DE TO AUTOCLAVI NG

P I ONE E RS I N S T E RI L I Z AT I ON T E CHNOL OGY

Sterilization may be defined as the statistically complete
destruction of all microorganisms including the most resistant
bacteria and spores. This is a condition that is difficult to
achieve and hard to prove. Whilst there are many chemicals,
inorganic and organic, that kill microorganisms they may not be
totally effective and can leave undesirable or toxic residues.
Ultraviolet and Ionising radiations are also effective biocides,
disrupting or modifying the DNA to prevent replication, but
Ultraviolet will not produce the effective results and easy
validation that moist heat (steam) sterilization can provide.
If sterility is an absolute requirement then todays scientists turn,
as their predecessors did, to steam.
Microorganisms tend to become more active as the
temperature of their surroundings rises, - most, but not all, die
at above 80
o
C. In the case of Prions the temperature and time
requirements for deactivation are much higher.
Steam molecules condense on cooler microorganisms, and
transfer 2500 joules per gram of steam, very efficiently heating
the microorganisms to the temperature at which they are
destroyed. Other methods of heating suffer from the much
lower heat transfer of hot dry gases and boundary layer effects,
which can insulate and protect the microorganisms.
For maximum effect the Steam must be saturated, and this
condition, and the temperature and pressure of the steam are
easily monitored, facilitating proof that sterilization has occurred.
By employing Steam Sterilization techniques
a high level of sterility can be achieved and
the most popular piece of equipment used
in laboratories and hospitals is the STEAM
STERILIZER or AUTOCLAVE.
The terms Autoclave and Sterilizer are interchangeable and mean
the same thing. Autoclave is a term used mainly in laboratories
whilst Sterilizer is used more commonly for medical and
pharmaceutical applications.
An effective Autoclave or Sterilizer must contain dry saturated
steam. In order to achieve this air must be removed from both
the load and the chamber; this can be accomplished in a number
of ways. Air removal from high mass low load items (i.e.
comprising mainly solid areas that contain little or no air pockets
such as bottled media) will need little air removal and this can be
facilitated by automatic air purging. Here air is allowed to leave
the chamber through a vent as steam enters the chamber either
from an integral source (upward displacement) or an external
supply (downward displacement); this vent only closes when all of
the air is removed from the chamber. This method can be further
advanced by Freesteaming when the vent is allowed to stay
open for a set length of time. The turbulent steam then passes
through the vent forcing any trapped air out of the autoclave. For
more stubborn loads that contain a number of air pockets such
as wrapped instruments or fabrics, a more effective method of air
removal is essential. By far the most effective way of air removal
is a vacuum system, whereby a vacuum is achieved in the
chamber before any introduction of steam, removing most of the
air before freesteaming and/or vacuum pulsing.
Once all of the air is removed from the load and chamber the
temperature within the vessel will rise along with the pressure until
the pre-selected temperature is reached. To achieve a typical
sterilization temperature of 121
o
C or more, requires the steam to
be pressurized to at least 1.1 Bar G. Since pressures used are
greater than 0.5 Bar G the Autoclave assembly is classified as a
pressure system and must be designed to a strict engineering
standard such as PED97/23EC/PD5500/ASME etc.
The temperature within the chamber is reached in a number
of ways:
1) Steam can be injected into the chamber via an internal
steam source such as an integral stainless steel steam
generator, which can be built within the Autoclave cabinet
or can be supplied as a separate unit.
2) Some laboratories or hospitals have their own direct steam
source, which would similarly be injected, into the chamber.
3) In some Autoclaves heaters are built into the base of the
chamber and the water is heated until it boils and produces
steam.
The main advantage of using a steam generator (or the direct
steam method) is that cycle times can be considerably faster as
the steam is immediately available. In basic autoclaves the
water is topped up manually by pouring water into the vessel
whilst some autoclaves are connected to a direct water supply
and the water level is maintained automatically. Some of Astells
smaller units are available with an integral water tank, which
once filled, can run for up to 20 cycles repeatedly before being
replenished. In CSSD applications it is a requirement that the
steam is dry saturated steam with a known dryness and non-
condensable gas content e.g. in the UK, Steam with values
within the specification set out in HTM2010.
When the desired temperature is achieved for the required time
then the steam supply will cease either by shutting off the power
to the heaters or by cutting off the steam supply. Thus the
temperature and the pressure will gradually drop. In units with a
vacuum system fitted the vacuum pump can be used to remove
the steam (i.e where drying is required).
How does an Autoclave work?
What is Sterilization?
Typical temperature/time sterilization parameters are 115
o
C for
30 minutes, 121
o
C for 15 minutes and 134
o
C for 3 minutes.
However, overall cycle times will vary depending on load
characteristics. Firstly it is important to realise that the time and
the amount of steam that is needed for the load to reach
Sterilization will vary greatly depending on the thermal capacity
of the load in question. Two or Three 1 litre bottles of culture
media will often need more steam than a dozen small
containers, and the heating time will take longer. Likewise the
time taken to achieve Sterilization once the temperature has
been attained will depend on the type of microbiological
organisms present. Bacteria and fungi are rapidly killed by moist
heat but viruses and particular bacterial spores are very
persistent, and may need long Sterilization times. The
development and wide availability of reliable microprocessors
means that most modern autoclaves are fully user
programmable, allowing the Sterilization temperature to be set
to suit the requirements of the load.
The most accurate way of ensuring that Sterilization occurs
when the correct temperature is reached is by Load Sensed
Process Timing. Here the cycle is controlled via the temperature
achieved in the centre of the load. A wandering thermocouple
probe situated within the chamber is inserted into the load, or a
load simulator and initiates the sterilization period once the
probe reaches the programmed threshold temperature. This
ensures that sterilization starts at the correct stage of the cycle.
Following Sterilization and depressurization of the chamber it
may be possible to open the autoclave immediately. However,
when autoclaving bottled fluids, a cooling lock is normally fitted
to comply with various safety regulations (e.g. in the UK: HSE
PM73). This prevents the autoclave from being opened until the
load temperature has fallen to typically 80
o
C, thus reducing the
risk of both the boiling over of fluids (e.g media) and the
explosion of glass bottles.
Following Sterilization it is possible to have a variety of fast
cooling options fitted which reduce the cool down period.
Deluge Cooling
This can often cut cycle times down by up to 50% and is used
only in conjunction with sealed fluid loads. Water at Sterilization
temperature is sprayed onto the load after the completion of the
sterilization stage. The water being sprayed is passed through a
heat exchanger cooled by cold water thus producing a
controlled reduction in temperature throughout the load. This is
a highly specialised method of cooling, and due to the volume
of water used during the deluge stage it is only suitable for
sealed bottled fluids.
Water Fed Jacket
A Jacket can be fitted to the autoclave chamber on both
circular and square section units. Again cold water is introduced
into the jacket at the end of the sterilization stage of the cycle
using the chamber walls as a heat exchanger reducing
temperature and resulting in improved cooling. As the jacket
acts as a very efficient heat exchanger pressure reduction can
be very fast using this method and therefore it is generally
recommended that Air Ballast is used in conjunction with the
jacket to reduce the possibility of fluid loss.
Advanced Water Cooling
This can only be fitted to machines that have been connected
to a mains water supply. A series of cooling coils are fitted to
the outside of the autoclave chamber. At the end of sterilization
these coils are circulated with cold water and so results in a
rapid decrease of temperature and improved cooling time.
Assisted Air-Cooling
Fans are fitted within the cabinet and circulate ambient air over
the external chamber walls during the cooling stage of the cycle
creating a controlled improvement to cooling times.
Internal Convection Fan Assisted Cooling
A fan or fans can be built within the chamber and ducted to
create an efficient airflow. The fan is again activated during the
cooling stage of the cycle. The fans create turbulence reducing
cooling time greatly. Internal convection fan cooling is one of
the most proficient ways of cooling the chamber. To ensure
improved cooling time the optional jacket and/or advanced
water-cooling is also needed. On loads using unsealed bottles
air ballast is recommended.
Typical Autoclave Cycles
Types of Cooling
Water Cooling
Fan Assisted/Air Cooling
The importance of drying!
It is important to take into account exactly what the autoclave
will be used for before running a cycle. Some items such as
fabrics & wrapped instruments require special attention as they
will need a specific drying cycle to avoid them being left wet at
the end of the cycle. The best way of ensuring that this does
not happen is by having a pre and post vacuum as well as a
steam heated jacket fitted to the chamber. The pre-vacuum is
required to remove air from the load and the post vacuum will
remove moisture left on the load. The drying will be enhanced
with the addition of the steam heated jacket, which is filled with
steam throughout the cycle; this heat prevents excessive
condensation being formed on the chamber walls and maintains
the temperature above the boiling point during drying so that
water droplets boil off. The Heated jacket is essential for
optimum drying performance.
Autoclaves can be used for numerous medical and laboratory
applications in Market Sectors such as, Food and Dairy,
Universities & Colleges, Agriculture & Horticulture Research,
Healthcare (including Pathology), Industrial Research, Water &
Environmental, Brewing, Biotechnology Industries and
Pharmaceuticals
Typical applications include:
Media Preparation
Sterilization of Fluids
Instruments (Wrapped and Unwrapped)
Utensils and Lab Equipment
Fabrics and Textiles
Media Preparation: Media is used to grow microorganisms
under test conditions and a wide variety of gel and liquid media
is used. Air removal from fluid/media loads is normally
straightforward; however some media cycles can benefit from a
short period of free steaming. Different types of media withstand
different sterilizing temperatures, therefore any autoclave used
for the Sterilization of media must be capable of sterilizing at a
variety of temperatures. Cycle times must be short and the
speed of processing is vital. A fast heat up and cool down is
beneficial in the maintenance of the quality of the finished
product and prevents the over cooking which leads to the
deterioration in the quality of the growth medium. Cycle length
should be set to as short as possible, and at the temperature
specified by the media manufacturer. The cool down period
however, needs to be a controlled process in order to prevent
boil over and loss of media. The main method of controlling the
sterilzation procedure of any cycle is to use a thermocouple
probe in a dummy bottle within the chamber (Load Sensed
Process Timing). Only when the probe reaches the correct
temperature will the sterilization process begin. Load Sensed
Process timing is used in conjunction with Profiled Overshoot
Boost, this allows the temperature in the chamber to rise above
the programmed level to allow the load to reach the correct
temperature faster. The risk of any boil over can be greatly
reduced by using Air Ballast which maintains chamber pressure
during the cooling phase.
Sealed Fluids: Some fluid loads may require to be sterilized in
special sealed containers. Air purging is normally sufficient to
remove air, and extended freesteaming should be unnecessary.
As with media preparation the speed of processing is vital to the
quality of the end product and often temperatures in excess of
121
o
C will be used to speed up the Sterilization process. Again
the cooling should be as short as possible.
Instruments:
The Sterilization
of instruments falls
into two main categories:
a) Unwrapped Instruments b) Wrapped Instruments
Both types of instrument loads benefit from drying the end of
the cycle. Unwrapped instruments can be dried by the heating
of the surrounding air at the end of the cycle, whilst wrapped
instruments require a more sophisticated drying system Post
vacuum systems on small bench-top units will improve drying,
whilst a jacketed pressure vessel is strongly recommended on
larger units.
Uses of Autoclaves
Ombar
1000mbar
2000mbar
3000mbar
START
VAC POSITIVE PULSE STERILIZE COOLING DRYING AIR BREAK
2100mbar
Ombar
1000mbar
2000mbar
3000mbar
START
VAC POSITIVE PULSE STERILIZE COOLING DRYING AIR BREAK
Ombar
1000mbar
2000mbar
3000mbar
START
VAC NEGATIVE PULSE POSITIVE PULSE STERILIZE DRYING AIR BREAK
Sealed Fluids Cycle Graph (including assisted cooling)
Typical Glassware/Utensils (134C) Cycle Graph
Sealed Fluids Cycle Graph
Fabrics and Textiles: A variety of fabrics and textiles can be
sterilized in an autoclave/sterilizer, e.g many hospitals will use an
autoclave to sterilise gowns, instrument packs etc. As these are
true porous loads they will require both pre and post vacuum
systems, negative and positive steam pulsing and a fully heated
jacket. Steam quality is important when operating Porous Load
Autoclaves.
Discard Loads: There is an ever-increasing need for the safe
Sterilization of contaminated material before disposal to ensure
that it is microbiologically safe. The contaminated material can
be of many types and in many forms. For example, all
microbiological laboratory waste should be processed prior to
disposal, as must all clinical samples e.g. blood, tissue etc.
Loads consisting of such a wide variety of materials in a broad
range of containers will present specific Sterilization problems.
One of the main problems with such mixed discard loads is
that they invariably contain small pockets of trapped air, which
present particular air removal problems before steam
penetration. Such loads are often contained in special plastic
bags and comprise a variety of plastic containers; these are
liable to melt and collapse before Sterilization , thus increasing
the likelihood of trapped air. A pre-vacuum and steam pulsing
feature is now a commonly requested option for autoclaves
running frequent discard cycles. However an alternative to
vacuum is a long period of freesteaming or Pulsed
Freesteaming
PAT
. There are special discard (Morrison) containers
available for this application. In certain circumstances a double-
doored autoclave may be required. The loading (dirty or non-
sterile) end is often separated from the unloading (clean or
sterile) end by a wall, which often contains an SPF seal to
prevent bacteriological migration. In category III laboratories an
effluent retention system would also be necessary.
Load Vacuum Assisted
Sensed Vacuum & Assisted Cooling
Process Pulsing Heated Cooling & Air
Timing Jacket Ballast
x x
(
3
)

x x x
x x
x x x x
x
(
1
)
x x
x
(
1
)
x x
x
(
2
)
x x
Ombar
1000mbar
2000mbar
3000mbar
START
VAC NEGATIVE PULSE POSITIVE PULSE STERILIZE DRYING AIR BREAK
Typical Fabrics Cycle Graph
Ombar
1000mbar
2000mbar
3000mbar
START
VAC POSITIVE PULSE STERILIZE DRYING AIR BREAK
COOLING
Typical Fluid Discard Cycle Graph
Media
Mixed
Discard
Fluid
Discard
Glassware
Unwrapped
Instruments
Wrapped
Instruments
Fabrics
Essential Recommended x Not Needed
(1) If dry load is required
(2) Vacuum & Heated Jacket required to ensure
drying of Fabric Loads
(3) To prevent excessive cycle time
Verification has become increasingly important, as
there is often a need to trace back through a series of
results to ensure that a particular Sterilzation process
has been carried out. There are a variety of options that
are available for this; these can either be mounted externally i.e
on an adjacent wall or shelf, or can be an integral part of the
autoclave. Printers, Chart Recorders or Dataloggers are the
preferred options. These instruments can provide either a hard
copy record of exactly what the microprocessor is registering, or
they can be completely independent from the microprocessor
and record independent results of chamber and load conditions.
Data Printers
Integral Data Printer
Built as an integral part of an autoclave this is the most popular
and cost effective way of monitoring the cycle. When the cycle
begins the printer records the starting time, cycle number,
program number (or name) and machine type. The printer will
print a complete record of cycle conditions, freesteam, sterilizing
start and finish time and pressure readings at one-minute
intervals. There is also provision for operator signature and
batch number.
Secureprint Data Printer
This is a Freestanding Data-
Printer independent of the
microprocessor, which is
designed to give an independent
printer verification of the sterilizing
function. Chamber temperature,
pressure, load and time is
displayed and printed with a dual
speed papersave feature.
(This printer will also operate with
any non Astell Autoclave)
Chart recorders
Chart recorders are the traditional method of printing the
Sterilization process. These show a graphical display of
pressure and/or temperature come with either one, two or three
pens, single pen recorders register chamber temperature, two
pen recorders register chamber temperature and load
temperature and the three pen recorders register chamber
temperature and load temperature via two wandering probes,
options also include measurement and display of pressure.
The recorders can either come in strip or circular format.
Circular Chart
Recorders
These are high
performance
microprocessor based
recorders with an
alphanumeric display
to provide more
information and easier
operation. The
recorders are available
with up to 1, 2 or 3
pens. Reading
accuracy is 1% of
Span of the
instrument. The
programme system allows full control of ranging, plus control of
the pen drive, lift and referencing, and display. The tactile
membrane switches are mounted at the front, alongside the
display, behind the door for security.
Strip Chart
Recorders
These are
microprocessor
controlled strip chart
recorders available
with 1,2 or 3 coloured
pens. Recording
accuracy is 1% of
span through the use
of accurate stepper
motors.
The recorder incorporates
a high definition
alphanumeric display, indicating temperatures and pressure.
Astell AutoSCRIBE Datalogger
The Astell datalogger is a purpose designed, wall mounted
datalogger with LCD display and keyboard.
The datalogger is specially configured to record temperature
and time parameters. The configuration of the AutoSCRIBE
allows maximum storage by only recording during the operation
cycle. The data is downloaded on demand to a laboratory
P.C. for analysis and archiving.
Monitoring the Sterilization process
Typical printout from integral or
Secureprint data printer.
(7)
(6)
(5)
(4)
(3)
The satisfactory performance of any Autoclave requires that it is
installed in accordance with the instructions supplied by the
manufacturer. Some small benchtop autoclaves require little or
no installation as they have an integral recirculation water tank;
therefore the machine requires no plumbing to a mains water
supply or drain. Where connection is required to the various
services (i.e water, drain, power etc) Astell supplies full
installation instructions and diagrams;
an example of which is shown.
Installation
It has become increasingly important to comply with Quality
Assurance procedures, which require validation that Sterilization
has been achieved. Validation is best defined as the
documented procedure of obtaining, recording and interpreting
results to ensure that the machine will consistently carry out a
defined process. This will involve carrying out multi-point
temperature mapping to establish suitable loading configurations
and cycle parameters to ensure that the process is controlled
within predetermined limits. Astell Scientific is a UKAS
accredited company.
For further details contact our Service Department.
1) Machine Power Switch
2) Power Supply= Switched FUSED connection unit
3) Chamber Exhaust carrying steam & condensate
4) Vent to external atmosphere or extraction hood
5) Connection to Drain- 35mm Copper. Must be
connected to drain with sealed plumbing
connection no gaps for steam to escape
6) Building Drain This must flow freely under gravity
away from machine;-Minimum diameter 50mm.
At no point must this slope uphill and there must
be no dead legs , loops or u-bends. Between
machine and the main drain system.
7) Chamber Drain for cleaning only. May be
plumbed to Building drain if required.
Validation
TEMPERATURE/PRESSURE CORRELATION
The table below shows Pressure and Temperature correlation for
Saturated Steam, and the Maximum/Minimum acceptable
Temperature display Values for a Correlation Accuracy of +/- 2%.
It applies to ASTELL Sterilizers only.
Pressure Temperature Temperature - Tolerance
Bar Deg C Min Temp Max temp
0.70 115.40 113.09 117.71
1.037 121.00 118.58 123.42
1.40 126.28 123.75 128.81
2.00 133.69 131.02 136.36
2.25 136.43 133.70 139.16
Typical installation (Electrically heated model)
(2)
(1)
Astell Scientific have been pioneers in the manufacture and
validation of Scientific and Laboratory equipment for over 80
years. In the last decade the company has concentrated on
becoming market leaders in Sterilization technology and is now
one of the premier manufacturers in this field in Europe.
A wide range of standard front and top loading microprocessor
controlled Autoclaves are offered including circular section units
from 23-330 Litres and Square Section Sterilizers from 100-490
litres. In recent years Astell has successfully developed its
bespoke manufacturing facility and offers a wide range of non
standard units in capacities up to 1000 litres. All of the units
feature the secure micro-computer controller, which is user
friendly and fully programmable. A variety of options and
accessories can be supplied with all models to ensure
optimum performance and user flexibility in the most
demanding situations. These range from Data Printers,
Load Sensed Process timing and a variety of cooling options
to reduce cycle time.
The Astell range can be catogorised as follows:
NB. For further information on Astells Autoclaves and Sterilizers
please refer to our separate brochure: The Autoclave Range
or visit our Website at www.astell.com
Astell - Pioneers in Sterilization Technology
Swiftlock Compact Autoclaves,
23, 40 & 60 Litres
This range of circular section autoclaves from Astell offer
guaranteed precision and versatility for a wide range of
applications. Featuring many of the benefits associated with
Astells larger units these stylish yet economical models meet
the demanding requirements of current Sterilization procedures.
These are now available in the Classic and Autofill versions.
Secure micro-computer control
316L stainless steel pressure vessels
Temperature range 100-138C (0.2 2.4 Bar)
Super-quick New Swiftlock door mechanism
Compact ergonomic designs
Automatic cycle control
Built in drying system (Autofill version)
Integral reservoir and/or condensate bottle
10 Year Pressure Vessel Integrity Warranty
Media holdwarm feature (Classic version)
Autoclaves, Swiftlock, 90 to 330 Litres
A range of top loading and front loading microprocessor
controlled autoclaves in volumes from 90 to 330 Litres. The
easy open/close Swiftlock closure combines with a user-friendly
control system to ensure simple operation for all applications,
whilst a comprehensive range of safety features ensures peace
of mind for the user.
Cost Effective
Full Heated Jacket Available for Optimum Drying
Secure Microcomputer control
PIN restricted user programming
Single action closure
Holdwarm for Media Preparation
Timed freesteaming
316L Stainless Steel Vessels
Cooling locks for the safe Sterilization of liquids
Single or multi-program models available
Temperature Range 100-138
o
C (0.2 2.4 Bar)
Optional Vacuum/AirBallast.
10 Year Pressure Vessel Integrity Warranty
Autoclaves, Square Section
(including models with Vacuum & Air Ballast)
A comprehensive range of Square and Rectangular Section
front-loading autoclaves, in standard volumes from 100 to 490
litres
Maximum Space Utilisation
Easy Loading
PIN restricted user programming
Fully interlocked sliding door.
316L Stainless Steel Vessels
Temperature range 100 138
o
C (0.2 2.4 Bar)
Standard sectional jacket with water cooling.
Optional Vacuum & Air Ballast
Double door units available
10 Year Pressure Vessel Integrity Warranty
The modular internal chamber dimensions of these units ensure
maximum space utilisation for all Sterilization purposes. The
models with vacuum facility are especially suitable for difficult
discard loads, ensuring efficient air removal prior to Sterilization.
All units feature the widely acclaimed Secure microprocessor
control system.
For further information on Astells Autoclaves and Sterilizers
please refer to our separate brochure: The Autoclave Range
or visit our Website at www.astell.com
Continuing efforts to improve products may necessitate alterations
to specifications without notice.
Astell Scientific Limited, Powerscroft Road, Sidcup, Kent DA14 5DT United Kingdom
T (020) 8300 4311 F (020) 8300 2247 E sales@astell.com www. astell.com
Direct Sales: T (020) 8309 2024/3/2 F (020) 8309 2036 0594 Certificate No: Q536