Systec VX 95 Series Manual

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 1 of 26
1. PURPOSE
To describe the procedure for the use and maintenance of the Systec VX 95 Series Autoclaves; No.
CBE045 (Don Whitley Serial No. 3099) and CBE044 ( Don Whitley Serial No. 3098) in the CBE
Autoclave Room (H31).
2. SCOPE
This SOP describes operational procedures for using the Systec VX95 Series (No CBE045/CBE044)
autoclaves for :
1)steam sterilization and safe disposal of solid biological waste that may contain biological agents
or Genetically Modified Organisms, as defined in the CBE Code of Practice, where solid waste may
include solid culture and stocks of infectious material, culture dishes, flasks and related devices,
sharps bins, contaminated solid items such as paper towels, plastic pipettes, pipette tips, vials and
gloves.
2) steam sterilization of aqueous solutions and media not exceeding 5L,
3)sterilization of reusable equipment (not waste) that may include bottles, beakers, pipettes, forceps;
tubing etc. All procedures described in this SOP for solid or liquid waste treatment have been
validated. Typical waste batches are described in Annexes at the end of the SOP.
SPECIAL NOTES: HEALTH & SAFETY
Risks to the operator
The autoclaves are state-of-the-art and built according to standard safety regulations. Nevertheless, the
danger of death or injury to the user or a third party, or damage to the autoclave or other material assets,
can arise when using the appliance. For this reason, the autoclave is only to be used as it is intended to
be used and in a faultless condition with regard to safety. NOTE: Faults that could impact on safety
must be resolved immediately. The following residual risks result from the function of the autoclave,
and must be considered at all times:
(i) Crushing and or amputation hazards - There is a danger of crushing or amputation of hands or
fingers between the edge of the door and the edge of the container if closing of the door is no
longer damped by spring or hydraulics.
(ii) Burn and scald hazards when unloading the autoclave (for some designated cycles) from (1)
hot vessels or liquid escape from vessels leaking (2) hot surfaces inside the container or near the
sterilisation substance. Hot surfaces of the door and chamber (3) escape of hot clouds of steam

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 2 of 26
on opening the door (4) unathorised or improper programing of parameters (5) unrecognised
errors and defects or poor maintenance procedures.
(iii) Heavy Load hazards - fully loaded baskets/buckets can be too heavy to lift manually when
putting them in or taking them out of the autoclave. If this is the case, the baskets or fillers should
be partially unloaded beforehand (max 35kg inclusive of basket or fillers).
(iv) Hazardous substances - Infectious aerosol hazards from (1) failed exhaust filter cartridges or
their improper disposal (2) non-sterile condensate left behind in the chamber if the sterilisation
process is not successfully completed
(v) Fire or explosion hazards from (1) misuse of sterilisation cycles ie sterilisation of liquids using
solid cycles (2) sterilisation of liquids in tightly closed containers (3) unauthorised or improper
programing of parameters.
(vi) Operational hazards - After using the autoclave, it must be ensured that the appliance is
properly switched off by means of the main switch (on the side of the autoclave)
(vii) Risks due to wear - The autoclave must be serviced at the regular intervals as specified in the
servicing regulations.( at least once a year or after 500 cycles).
(viii) Specific risks of individual sterilization procedures - Additional specific risks can arise when
using particular sterilization cycles. Specific instructions regarding these risks and how to avoid
are detailed in the operating manual.
(ix) Incorrect use of the cycles for solids - When sterilizing liquids in glass containers with the
cycle for solids, the boiling process can be delayed and the glass container can shatter.
Safe Working Practices
In addition to the basic and specific safety instructions, risks can arise through dangerous working
procedures. NOTE: Detailed information on laboratory autoclaves can be found in British Standard
BS2646 ‘Autoclaves for sterilisation in laboratories’, in the HSE publication PM73, ‘Safety at
autoclaves’ and in the local Code of Practice.
(i) The operator MUST ensure that the item to be sterilized is suitable for steam sterilization
in the autoclave with the options installed.
(ii) The operator MUST ensure that the maximum load capacity for the autoclave is never
exceeded (refer to Operators Manual for specifications).
(iii) Hazardous chemicals or materials contaminated with them MUST NOT be autoclaved.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 3 of 26
(iv) Materials contaminated with Virkon are NOT to be autoclaved.
(v) Any strong oxidizing material i.e. dry hypo chlorite’s, MUST NOT be autoclaved with
organic materials such as paper, cloth or oil.
(vi) While contaminated solid biohazardous waste, reusable equipment or aqueous solutions
may be sterilized in the same autoclave, they MUST NOT be mixed or loaded together during
the same cycle.
(vii) The main switch switches the power supply to the autoclave on and off. ALWAYS switch
off the autoclave at the main switch after use, or if some danger has been detected in the
autoclave.
(viii) The preset cycles of the autoclave can be changed significantly using the operational
parameters described in the manual, and this may result in danger for the operating personnel
or impair the sterilization result. Changes MUST only be made by trained personnel and MUST
be documented in the appliance log book.
3. RESPONSIBILITES
CBE Laboratory Users
(i) Only persons who have received the appropriate training are permitted to use
autoclaves. The names of the authorised users should be posted on or nearby the autoclave.
(ii) All operators MUST observe all safety regulations and guidelines applying to the
autoclave and the environment in which it is operated
(iii) All operators MUST read the Risk Assessment and this SOP and confirm with their
signature that they have understood both documents. A signed Training Agreement should be
retained in the individual’s Training Record.
(iv) All operators MUST use the prescribed PPE; heat resistant gloves, eye protection and
an impervious apron (SOP037)
(v) All operators MUST keep the autoclave in a faultless condition with regard to safety.
(vi) All operators MUST stop using the autoclave as soon as any safety deficiency is
detected. Operators MUST inform the Laboratory Manager or designated person. All error
messages must be reported.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 4 of 26
(vii) Autoclaved items should be stored separately from non-sterilised items. There should be
designated laboratory areas for storage of items related to their contamination status and prior /
intended use e.g. clean equipment for autoclaving, autoclaved equipment, and -contaminated
used equipment for cleaning, contaminated used equipment for disinfection. All laboratory users
should be aware of the system.
(viii) Authorised users should check that the autoclave is in service and has been validated
for use. Autoclaves are inspected annually and certified by the service engineer or designate.
The inspection, service and repair records are maintained as per University Safety Policy. ‘Next
service’ date and ‘next validation’ date should be posted on / near the autoclave.
(ix) All users must check the cycle has worked correctly by checking the screen and
printout.
(x) All users must complete the log for each load.
Responsible Person (RP)/Laboratory Manager (LM)
(i) The Laboratory Manager or designated Responsible Person MUST ensure that the University
Estates and Buildings Department is notified of all autoclave installation. They will inspect the
autoclave at the statutorily required interval. Notification of such items should be made through
the Departmental Safety Officer (DSO) and the Health and Safety Department.
(ii) Notification of newly acquired equipment is required before it is brought into use, to ensure
compliance with the Pressure Systems Regulations.
(iii) The Laboratory Manager or designated Responsible Person is responsible for ensuring that the
autoclave is validated for the type of waste generated in the CBE Laboratory Unit. Annexes list
the typical waste loads for which the autoclave is validated for at the time of writing. If typical
waste loads differs significantly from those validated, personnel MUST inform the Laboratory
Manager so that arrangements can be made to validate the prospective new loads.
(iv) The Laboratory Manager or designated Responsible Person is responsible for checking the
usage log to ensure that the autoclave is serviced correctly; either on its annual ‘due date’ or
when it has reached its 500 cycle limit.
(v) The Systec VX series autoclave pressure vessel is serviced annually by the Systec engineers
as part of the service contract. It is the Laboratory Managers or designated Responsible Person
duty to ensure that the Insurance inspection is performed with the Systec engineer present and
the inspection certificate is lodged with Technical Resources Manager in the Wolfson School.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 5 of 26
Mastercopy
(vi) Principal investigators/supervisor’s MUST ensure that his/her employees are trained before
operating any autoclave unit. All operators (regardless of experience) must have successfully
completed an authorized training session on the safe operating procedures of autoclaves.
Users must be instructed on the operation and safety regulations and, if required, to repeat this
instruction at regular intervals
(vii) Maintenance of autoclaves should not commence without a 'Decontamination Certificate'.
4. EQUIPMENT AND MATERIALS
1) Systec VX-95 Autoclaves Number CBE045 and CBE 044
2)Printer
3)De-ionised water vessel
4)Autoclave Basket
5)Autoclave Bucket –item J807 from nisbets.co.uk
6)Integrated hoist
7)Autoclave Log Sheets
8)Autoclave bags (Fisherbrand, polypropylene or HDPE)
9)Autoclave Indicator Tape
10)Polypropylene or stainless steels tubs as secondary containment
11)Sharpsafe one Sharps autoclavable containers (nominal volume –1L),
5. PROCEDURE
The following describes procedures for (i) the validated steam sterilization of solid form biological waste
that may contain biological agents or Genetically Modified Organisms. (ii) The sterilization of solids,
reusable equipment and (iii) the sterilization of liquids, aqueous solutions and media.
In addition to this SOP and the Operators manual, simple, easy to understand instructions are
provided for the operator in the autoclave room.
NOTE; before proceeding all operators must don
oFull length apron (green)
oEye protection (safety goggles or full face visor)
oSpecialised Heat Resistant (Orange) Gloves

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 6 of 26
5.1 Opening the Autoclave Door
(i) Turn on the power for the autoclave
(ii) CAUTION: Before opening the door check the log to see if the autoclave has recently
performed a sterilisation cycle. If so the surfaces of the door and the autoclave chamber will be
hot and on opening, hot clouds of steam can escape and lead to scalding.
(iii) To open the door, press the “Open” button displayed on the front panel. The autoclave will
alarm for 30 seconds to warn the operator to move back. The door is unlocked and
automatically opens slightly. The operator should stand clear and wait 1 minute before opening
the door fully.
(iv) Wearing the appropriate PPE, lift the door until it reaches the vertical stop. DO NOT pull the
door from the front,
(v) Ensure that the flexible temperature sensor (PT100) is placed in the holder in the door when the
door is opened. HINT: It is a good idea to put it in a coil, so that it does not trap in the seal of the
autoclave.
(vi) NOTE: Before using the autoclave, ensure that the distilled water reservoir is filled to the
marked level.
5.2. Selecting the Sterilisation Cycle
Depending on the item being sterilized, you MUST select an appropriate sterilization procedure in order
to:
•eliminate risks to personnel and to the laboratory
•Guarantee a successful sterilization result.
A suitable sterilization procedure is selected using the arrow keys on the control unit. The corresponding
cycle number, the name of the cycle and the pictogram for the temperature sensor used appears in the
display. Eleven sterilization cycles, one cleaning cycle and two test cycles are pre-programmed in the
factory setting.
Only three of these cycles are authorised for routine sterilisation as detailed in the following
sections of this SOP (see Table 1). The use of the other cycles MUST NOT be used unless
authorised by the Laboratory Manager or Responsible Person.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 7 of 26
ALL waste must be sterilised using validate cycles. Non validated cycles should only be used for
sterilising equipment which will remain in the laboratory.
Table 1: Preset Parameters for Sterilisation Cycles
Item being Sterilized
Sterilization
Temp
Sterilization
Time
Unloading
Temp
Cycle No.
In Combination
with:
Validated
(Yes/No)
Solids, instruments
121 °C
20 min.
120 °C
1
N/A
NO
Solid Laboratory waste
(non melting items) in
Bags in the basket
121 °C
15 min.
99°C
4
N/A
YES
Solid Laboratory waste
(melting items) in Bags
in steel bucket in basket
121 °C
15 min.
99°C
5
N/A
YES
Liquid Laboratory waste
in bottles (max 5L with
probe in 2L reference
bottle)
121 °C
15 min.
80°C
6
N/A
YES
Liquids in containers
121 °C
15 min.
80 °C
11
N/A
NO
Liquids in containers
121 °C
20 min.
80°C
8-10
Rapid Cooling
NO
CAUTION: While contaminated solid biohazardous waste, reusable equipment or liquid may be sterilized
in the same autoclave, DO NOT mix them together during the same cycle.
5.2.1. Cycle Selection
(i) CYCLE 1 (Solids, instruments) must be used for steam sterilisation of solid equipment
at
121°C for 20 min.
(ii) CYCLE 4 (Solid waste in bags) MUST be used for the sterilization of normal laboratory
waste in bags (solid waste with low liquid content). The operating cycle, fixed at 121°C and
20 minutes, MUST be used for sterilisation of all solid biohazardous waste. This cycle has
been validated for worst case loads (a maximum amount of typical solid biohazard wastes)

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 8 of 26
based on an overkill sterilisation cycle approach (refer to Reference 4). Cycle 4 was
validated for 4 bags containing typical loads of solid waste (described in Annex 1).
(iii) CYCLE 5 (Solid waste with a potential to melt and Sharps bins) MUST be used for
the sterilization of solid laboratory waste that (a) has the potential to melt or has a high liquid
content eg agar plates - MUST be placed in bags and contained in secondary steel buckets
when autoclaved; or (b) autoclavable plastic sharps bins –MUST be placed in steel buckets
when autoclaved. The operating cycle, fixed at 121°C and 20 minutes, MUST be used for
sterilisation of all solid biohazardous waste. This cycle has been validated for worst case
loads (a maximum amount of typical solid biohazard wastes) based on an overkill
sterilisation cycle approach described in Annex 2 .
(iv) CYCLE 6 (liquid waste –up to a maximum of 5L). This cycle MUST be used for
sterilisation of liquid waste. Liquid waste must be kept in an autoclavable container. The
container MUST NOT be tightly sealed. A Reference volume (blank) containing 2L of
distilled water must be kept open (described in Annex 3). This is as per the validation
Certificate provided by Don Whitley Scientific Limited. A copy is available in the Systec VX-
95 equipment folder. The load-probe must be placed in the blank bottle.
(v) CYCLES 8-10 (Liquids in containers) must be used for liquids in autoclavable containers
when rapid cooling using mains water and compressed air is required.
(vi) CYCLE 11 (Liquids in containers) must be used for liquids in autoclavable containers
when rapid cooling using mains water and compressed air is not required.
NOTE: All waste material must be autoclaved using the appropriate one of the three
validated cycles (cycles 4, 5 or 6). If in doubt consult the Laboratory Manager or
Responsible Person.
5.3. Loading the Autoclave
5.3.1 Sterilisation & Disposal of Solid Form Waste with low liquid content and low propensity
to melt: CYCLE 4
Waste for this cycle includes solid form waste which has low propensity to melt or has a low liquid
content. Examples of such waste is given in Annex 1.
(i) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal Protective
Equipment (PPE).
(ii) Use CBE approved transparent autoclave bags. DO NOT use any other bags!

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 9 of 26
(iii) Label the bags to identify the laboratory area from which it came
(iv) Ensure that the solid form waste is free from excess liquids and sharp objects that could
puncture the waste bag. CAUTION: Sharps MUST be autoclaved separately using Cycle 5
(v) Do not fill bags more than ½ full. DO NOT seal the bags. If the tops are taped or tied
they MUST be opened loosely to ensure effective steam penetration.
(vi) Place bags in a secondary container –Use the wire mesh baskets provided! Wire mesh
baskets may prevent the autoclave becoming dirty or damaged due to leaks in rubbish bags or
other containers. Examine the secondary container for damage before using. DO NOT use any
secondary container other than those designated for the purpose.
NOTE: Fully loaded tubs/baskets can be too heavy to lift manually! To assist with the
loading and unloading of the autoclave, an electrically-operated lifting hoist is available (Max
load = 35Kg). It is mounted directly onto the appliance. It enables you to load and unload heavy
loads without any exertion. A swivel arm also facilitates operation.
(vii) DO NOT place the flexible temperature sensor in the item being autoclaved (ie the item
may melt!). The temperature sensor should be placed in the holder provided for it in the
sterilization chamber, or else left freely in the chamber. The coil for the probe must be
positioned so that it does not block the seal.
(viii) DO NOT overload the autoclave. Overloading the autoclave can lead to insufficient
ventilation of the autoclave and may cause faults to occur. For an optimal result, the steam
must reach all parts of the item being sterilized.
(ix) Place autoclave indicator tape (white banded tape) on outside of the autoclave bag.
(x) DO NOT leave bags for autoclaving in the autoclave overnight. Autoclave the load as
soon as possible after preparation.
5.3.2 Sterilisation & Disposal of Solid Form Waste with high liquid content or high propensity
to melt :CYCLE 5
Waste for this cycle includes solid form waste which has a propensity to melt or has a high liquid
content. Examples of such waste is given in Annex 2.
(i) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal Protective
Equipment (PPE).

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 10 of 26
(ii) Place the waste in CBE approved transparent autoclave bags. DO NOT use any other
bags!
(iii) Label the bags to identify the laboratory area from which it came
(iv) DO NOT seal the bags. If the tops are taped or tied they MUST be opened loosely to
ensure effective steam penetration.
(v) Place bags in the steel bucket described in the Equipment and Materials List in Section
6. Then place the buckets in the wire mesh baskets provided. Examine the secondary container
for damage before using. DO NOT use any secondary container other than those designated
for the purpose.
NOTE: Fully loaded tubs/baskets can be too heavy to lift manually! To assist with the
loading and unloading of the autoclave, an electrically-operated lifting device is available (Max
load = 35Kg). It is mounted directly onto the appliance. It enables you to load and unload heavy
loads without any exertion. A swivel arm also facilitates operation.
(vi) DO NOT place the flexible temperature sensor in the item being autoclaved. The
temperature sensor should be placed in the holder provided for it in the sterilization chamber, or
else freely in the chamber. The coil for the probe must be positioned so that it does not block
the seal.
(vii) DO NOT overload the autoclave. Overloading the autoclave can lead to insufficient
ventilation of the autoclave and may cause faults to occur. For an optimal result, the steam
must reach all parts of the item being sterilized.
(viii) Place autoclave indicator tape (white banded tape) on outside of the autoclave bag.
(ix) DO NOT leave bags for autoclaving in the autoclave overnight. Autoclave the load as
soon as possible after preparation.
5.3.3. Sterilisation & Disposal of Sharps Bins: CYCLE 5
(i) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal Protective
Equipment (PPE).
(ii) CAUTION: Sharps MUST be placed in the approved, autoclavable sharps bins
provided. Autoclave tape should be placed across the top of the sharps box.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 11 of 26
(iii) Seal the sharps bin following the instructions on the side of the container
(iv) Place sharps boxes in the steel bucket described in the Equipment and Materials List .
Then place the buckets in the wire mesh baskets provided. Examine the secondary container
for damage before using. DO NOT use any secondary container other than those designated
for the purpose.
NOTE: Fully loaded tubs/baskets can be too heavy to lift manually! To assist with the
loading and unloading of the autoclave, an electrically-operated lifting device is available (Max
load = 35Kg). It is mounted directly onto the appliance. It enables you to load and unload heavy
loads without any exertion. A swivel arm also facilitates operation.
(v) DO NOT place the flexible temperature sensor in the item being autoclaved (ie the item
may melt!). The temperature sensor should be placed in the holder provided for it in the
sterilization chamber, or else freely in the chamber. The coil for the probe must be positioned so
that it does not block the seal.
(vi) DO NOT overload the autoclave. Overloading the autoclave can lead to insufficient
ventilation of the autoclave and may cause faults to occur. For an optimal result, the steam
must reach all parts of the item being sterilized.
(vii) After autoclaving is over sharps bins must be removed and stored in the “Autoclaved
Sharps bin” box in the Waste Cage in H31 before being transferred to Gas Pod 2.
5.3.4. Sterilisation and Disposal of Liquid Waste (CYCLE 6)
Waste for this cycle includes liquid waste (maximum of 5L) in autoclavable containers.
(i) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal Protective
Equipment (PPE).
(ii) Place the liquid to be sterilised in autoclavable containers and place in the wire basket.
DO NOT tighten the container lid. Place the basket in the bottom of the autoclave. Ensure that
the liquid being sterilised is less than or equal to 5L in volume.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 12 of 26
Mastercopy
(iii) Prepare a second container with distilled water (the Blank). This must equal the volume
of the liquid to be sterilised to a maximum of 2L. Place the second container in a second wire
basket and place on top of the basket containing the item to be sterilised.
(iv) Place the load-probe in the distilled water and leave open.
(v) Label the bottles to identify the laboratory area from which they came
NOTE: Fully loaded baskets can be too heavy to lift manually! To assist with the loading
and unloading of the autoclave, an electrically-operated lifting device is available (Max load =
35Kg). It is mounted directly onto the appliance. It enables you to load and unload heavy loads
without any exertion. A swivel arm also facilitates operation.
(vi) Place autoclave indicator tape (white banded tape) on outside of the bottle.
(vii) DO NOT leave bottles for autoclaving in the autoclave overnight. Autoclave the load as
soon as possible after preparation.
5.3.5. Sterilisation of solid form reusable equipment, instruments and vessels- Non-validated
Cycles)
5.3.5.1. Cycle Selection
(i) CYCLE 1 (Solids) MUST be used for the sterilization of all kinds of solids, such
as instruments, glass and other materials for which the manufacturer recommends
sterilization in the autoclave. CAUTION: Make sure that the solid items such as
reusable equipment are suitable for autoclaving.
(ii) The operating cycle fixed at 121°C and 20 minutes MUST be used for
sterilisation of
all solids (except solid waste in bags). This cycle has NOT been validated. NOTE: If
solid items are to be used in critical or costly experimental procedures it is
recommended that this cycle is validated
5.3.5.2. Loading the autoclave with solid form reusable items: Starting the Cycle
(i) CAUTION: Cycle 1 must never be used for liquid waste.
(ii) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal
Protective Equipment (PPE).

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 13 of 26
Mastercopy
(iii) Ensure that all solid items are suitable for steam sterilization at temperatures
>121 °C or >134 °C and ensure that any packaging used is permeable to steam.
(iv) Inspect all solid vessels and other items for damage. DO NOT autoclave
damaged items; consult the Laboratory Manager or Responsible Person.
(v) Place the items either on the perforated bottom plate or in the designated wire-
mesh
basket. DO NOT use tubs or similar vessels –the items could be damp or even wet
when being taken out. DO NOT use any secondary container other than those
designated for the purpose.
NOTE: Fully loaded tubs/baskets can be too heavy to lift manually! To assist with
the loading and unloading of the autoclave, an electrically-operated lifting device is
available (Max load = 35Kg). It is mounted directly onto the appliance. It enables you to
load and unload heavy loads without any exertion. A swivel arm also facilitates
operation.
(vi) Label the bags or secondary containers to identify the user and laboratory area
from which it came.
(vii) Cap medium/solution bottles loosely or cover with aluminium foil. CAUTION: DO
NOT place sealed containers in the autoclave. Tightly capped medium/solution bottles
may explode due to expansion.
(viii) Cover bottles that are not made of safety glass with aluminium foil.
(ix) Place small items such as scissors, forceps, tubing, tube connectors into an
autoclave paper bag.
(x) Place autoclave tape on the top of the free-standing vessels such as glass ware,
pipette tip boxes or the paper bags and label it with your name.
(xi) Place the flexible temperature probe in the holder provided for it on the lid. The
coil for the probe must be positioned so that it does not block the seal.
5.3.6. Sterilisation of Liquids in open vessels (NOT liquid waste!) –Non-validated cycles
NOTE: The sterilisation of liquid waste in bottles or vessels has NOT been validated. If liquid waste is
generated, users MUST consult the Laboratory Manager or Responsible Person before proceeding.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 14 of 26
Mastercopy
5.3.6.1. Cycle Selection Only
use these cycles for liquids!
WARNING: The sterilization of liquids in solids cycles is highly dangerous through boiling
delays or exploding vessels and can result in serious injuries. For this reason, liquids may only
be sterilized using the cycle Nos. 6 to 11, which are designed for this purpose!
Liquids MUST be sterilized in OPEN vessels. For the sterilization of OPEN vessels you can use one of
the cycles with water cooling (Nos. 8, 9 or 10) as well as the cycle without active cooling (No. 11).
Liquids in hermetically closed vessels can be sterilised using the option for rapid cooling with support
pressure supply (No's 8, 9, and 10). If the sterilisation of liquid in CLOSED vessels is required users
MUST first consult the Responsible Person.
NOTE: These cycles have NOT been validated. If liquid items are to be used in critical or costly
experimental procedures it is recommended that this cycle is validated.
(i) CYCLE 11 (Liquids), with an operating cycle fixed at 121°C and 15 minutes,
MUST be used for the sterilisation of liquids in suitable OPEN vessels, without active
cooling. DO NOT USE FOR STERILISATION OF LIQUIDS IN TIGHTLY-SHUT
VESSELS!
(ii) CYCLES 8-10 (Liquids) should be used for sterilisation of heat sensitive liquids
in suitable OPEN or HERMETICALLY CLOSED vessels only. By means of rapid
cooling with cooling water with support pressure by filtered compressed air, the cooling
periods are reduced by approx. 70% in comparison to normal cooling. The loss of liquid
in the item being sterilized during rapid cooling with cooling water and support pressure
amounts to < 1%.
The operator can set a cooling rate that fixes a minimum cooling time, depending on the
cooling system and the loading. The minimum cooling time ensures that, regardless of
the unloading temperature, the autoclave can only be opened when this cooling time
has elapsed. The cooling rate (K/min) must be determined and set by the user,
depending on the type of load.
WARNING: With closed containers, water cooling may only be used in conjunction with
support pressure supply. If no support pressure supply is available, the containers must
be open!
(iii) The autoclave is fitted with a temperature dependent door lock. For sterilisation
of liquids, the flexible temperature sensor MUST therefore be placed in the liquid or in a

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 15 of 26
reference vessel. The reference vessel should have the same size and fill volume as the
vessel containing the liquid to be sterilized.
5.3.6.2. Loading the autoclave with vessels containing liquids (Cycle 8-11)
(i) ALWAYS wear appropriate PPE as described in SOP 037, Use of Personal
Protective Equipment (PPE).
(ii) Use temperature-resistant vessels! For the sterilization of liquids, it is
absolutely necessary to select temperature-resistant vessels that can withstand the
sterilization temperatures (>121 °C).
(iii) Make sure that the vessel is suitable for use with the selected cycle.
•If using Cycle 11 ensure that the vessels to be sterilized are OPEN - under no
circumstances vessels should be tightly-shut! Cap medium/solution bottles loosely –
whether empty or filled –to prevent explosions due to expansion.
•If using Cycles 8, 9 or 10: if vessels are to be sterilised CLOSED, ensure that
vessels are specifically designed to be hermetically sealed.
(iv) Place the vessels filled with liquid on the perforated bottom plate or in the wire-
mesh basket. DO NOT use tubs. DO NOT use any secondary container other than
those designated for the purpose.
(v) DO NOT overfill the open vessels - the filling height of open vessels should less
than 75% of the fill volume.
(vi) Large volumes of liquid will heat up (and cool down) slowly, therefore volumes of
liquid should be kept small; a maximum volume of 500 mL is recommended.
(vii) DO NOT autoclave solutions containing high concentrations of volatile or
corrosive chemicals. Seek advice from the Laboratory Manager.
(viii) Label the vessels or basket to identify the user and the laboratory area from
which it came
NOTE: With open vessels, boiling can reduce the volume of the liquid to be sterilized
after the sterilization process by up to a maximum of 5%.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 16 of 26
(ix) Place the flexible temperature sensor in the liquid or in a reference vessel. The
reference vessel should have the same size and fill volume as the vessel containing the
liquid to be sterilized.
5.4. Closing the Autoclave Door
(i) Once the autoclave has been loaded in accordance with instructions, close the
autoclave door by holding firmly shut until the door lock engages.
(ii) The ‘Ready’ message in the display tells you that the door is completely locked.
5.5. Starting the cycle
Make sure there is sufficient paper in the printer!
(i) Once the door is securely locked, select the correct cycle by pressing the menu key.
When the menu displays “Select Cycle” press the “Set” key. The list of cycles will then be
displayed. Choose Cycle 1, 4 or 11 using the arrow keys and then press the “Set” key.
(ii) In the solids cycles (1, 2 and 3), no liquids may be sterilized. The sterilization of liquids
in solids cycles causes danger through boiling delays or exploding vessels and can result in
serious injuries. To prevent liquids mistakenly being autoclaved in a solids cycle, a safety query
is performed after the Start display key is pressed. The message ‘No Liquids!!!’ informs you
that the cycle is not suitable for the sterilization of liquids. When you are sure that no liquids are
in the autoclave, confirm the query by entering the code of your access level.
NOTE: Liquids MUST only be sterilized using the cycle No's. 6 to 11, which are designed for
this purpose.
(iii) From this point on, the controller takes over the entire procedure. In the display you will
see a plain text report about the first phase of the sterilization program. NOTE: The printer
automatically logs the entire cycle procedure from the start of the cycle.
(iv) Preselectable starting time: If the parameter StartByTime is set to “1” in the cycle
selected, the current time and the current date appear after the Start display button is pressed.
You can use the arrow keys to set the desired start time and press the Start display button to
enter this time. Instead of the current time, the time remaining until the start is displayed.
5.6. End of cycle
After the completion of an autoclave cycle, an acoustic signal sounds and the message ‘Cycle ended’
appears in the display. The autoclave procedure is completed and the item being sterilized can be taken

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 17 of 26
out and used in accordance with the prescribed procedure, as described in the sections “Opening the
door” and “Taking out the autoclaved item”.
5.6.1. Stopping the cycle
If you wish to end a cycle prematurely, you must stop the procedure in progress.
•Press the Stop display key. Display message ‘Manual Stop’ after user stops cycle
•Confirm the error message by pressing the Quit display key and entering the code for the
access level. After a manual stop, the interrupted autoclave process cannot be continued, but must
be started again if desired.
(i) Items possibly not completely sterilized!
•Depending on the point at which the sterilization process is interrupted, the sterilization
may be incomplete. An interruption in the warm-up or sterilization phase causes the cycle to
pass directly to the cool-down phase. In this case, to guarantee complete sterilization you
must repeat the entire sterilization process!
•By interrupting the process in the cooling-down phase, for example, you do not speed
up the process. Please consider that the door can only be opened when the unloading
temperature has been reached and atmospheric conditions prevail inside the chamber.
(ii) Cycle error
If an error occurs, the “ERROR” LED lights up and a corresponding error message appears in
the display.
•Press the Quit display button and enter the code for the corresponding access level.
•Press the Set display key to confirm the error message.
(iii) Items possibly not completely sterilized!
Depending on the point at which the sterilization process is interrupted, the sterilization may be
incomplete. An interruption in the warm-up or sterilization phase causes the cycle to pass
directly to the cool-down phase. In this case, to guarantee complete sterilization you must
repeat the entire sterilization process!
Meaning of the error messages!
If you are not clear about the meaning of an error message and how to resolve the problem,
consult chapter 8: “Description of errors” of the Equipment manual.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 18 of 26
Mastercopy
5.7. Opening the door
(i) When the sterilization process is completely finished and the specified unloading
temperature and atmospheric conditions have been reached, the message ‘Opening Door’
appears in the display. The door can now be opened see (ii).
NOTE: If the sterilization process is stopped prematurely, the error message ‘Manual Stop Fail’
appears in the display. After you press the Quit display button and enter the code for the
corresponding access level, the door can be opened (after the unloading temperature and
atmospheric conditions set have been reached).
(ii) CAUTION: Before opening the door check the log to see if the autoclave has recently
performed a sterilisation cycle. If so the surfaces of the door and the autoclave chamber will be
hot and on opening, hot clouds of steam can escape and lead to scalding.
(iii) To open the door, press the “Open” button displayed on the front panel. The autoclave
will alarm
for 30 seconds to warn the operator to move back. The operator should stand clear. The door is
unlocked and automatically opens slightly. The operator should stand clear and wait 1 minute
before opening the door fully.
(iv) Wearing the appropriate PPE, lift the door using the grip handles on the side until it
reaches the vertical stop. DO NOT pull the door from the front.
(v) Ensure that the flexible temperature sensor (PT100) is placed in the holder in the door
when the door is opened.
5.8. Unloading the Autoclave: Taking out the autoclaved item
The door must be completely open!
When the autoclave procedure has ended and the once door has been opened, adhere to all safety
precautions when removing an autoclaved item from the autoclave (Refer to the special notes on health
and safety; section 4). The following steps must be taken:
(i) If the load inside the autoclave is not yours, check the log and ask the owner to remove
the load. DO NOT remove a load with which you are unfamiliar with its contents.
(ii) ALWAYS wear appropriate protective clothing when unloading the autoclave to avoid
being burned or scalded.
CAUTION: On opening the autoclave, tightly-closed vessels can still be at temperatures above
the unloading temperature.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 19 of 26
Mastercopy
CAUTION: There is danger of crushing and or amputation between the door and the edge of the
autoclave chamber if the door has not been completely opened.
(iii) If required, remove the flexible temperature sensor from the autoclaved item (or
reference vessel) and fix it to the holder provided for it in the door.
(iv) If fully loaded tubs/baskets are too heavy to lift manually they should be partially unloaded
before removal from the autoclave. Alternatively an electrically-operated lifting device is available
(Max load = 35Kg) to assist unloading. It is mounted directly onto the appliance. It enables you to
load and unload heavy loads without any exertion. A swivel arm also facilitates operation.
(v) Remove the basket, tub or item from the autoclave chamber and check the indicator tape:
➢Totally black lines indicates that steam exposure was effective
➢Not totally black lines indicate failure during the discard cycle. Re-autoclave the waste
by loading with sterilisation indicators and record the results in the User Log ➢If it is fails
again, report the problem to the responsible person or laboratory manager
5.9. Switching off the autoclave
(i) After use, switch off the autoclave at the main switch located at the side of the autoclave
(ii) If some danger has been detected in the autoclave switch off the autoclave at the main
switch.
NOTE: When the power supply is switched off during operation, the current process is
interrupted. The autoclave gradually returns to a depressurized state and low temperature, and
can then be opened safely. After the power supply is interrupted and then switched on again,
the autoclave returns to the state it was in before the power supply was interrupted. Any error
messages that may have been displayed on the control panel before the interruption are
displayed exactly as before. The pressure and temperature values displayed reflect the current
state.
(iii) In case of emergency; to shut off the steam source switch the power off using the
emergency safety switch located on the ceiling of the autoclave room above the autoclaves
(NOTE: a pole is provided to enable the operator to perform this action). Leave the autoclave
room and prevent others from entering. Consult the Laboratory Manager or Responsible Person
and contact the DSO.

CONTROLLED COPY
Centre for Biological Engineering
Standard Operating Procedure SOP025
Title: USE AND MAINTENANCE OF THE SYSTEC VX 95 AUTOCLAVES
Location: Autoclave Room, H31, CBE
Version 004 Effective Date: 28th June 2020 Review 28th June 2022
Written by: A. Chandra
Reviewed by: P.Hourd
Approved by: R.I.Temple
Page 20 of 26
5.10. Autoclave Load Testing and Recording
Autoclave runs used for waste management must be checked on each occasion by the use of recording
strips, data logging, etc. and maintained so as to ensure effective functioning and data recording. There
is a need to demonstrate reliable inactivation of waste by validation. Although autoclaves undergo annual
performance and safety checks, autoclave cycles may be inconsistent due to the diversity of loads placed
within them. Autoclaves which have a load temperature probe and printout of cycle parameters effectively
monitor the sterilisation process.
(i) The operator MUST record details of each cycle run in the User Log each time the
autoclave is used. This should include the following:
•Basic details such as operator name, date of use, cycle number should be recorded on
the log sheet provided
•The printed record that documents parameters while a cycle is running and provides
cycle data at the end of the cycle MUST be detached and the Log number from the template
form written on the printed record. The printed records MUST be signed and retained in the
Equipment File.
(ii) All printed records of cycle conditions and tests MUST be retained for at least 5
years.
(iii) Records must also be kept of all maintenance and service activities, reporting of
incidents, accidents and/or faults. All failed tests must be reported to the Laboratory Manager.
(iv) The autoclave must be inspected annually and certified by the service engineer or
designate.
The inspection, service and repair records should be maintained in the Maintenance & Service
Log .
5.11. Validation of Autoclave Performance
The purpose of validation is to test the ability of the autoclave to effectively decontaminate/sterilise
waste or wrapped/unwrapped goods when loaded with user-defined loads. Validation should
demonstrate that in the holding part of the cycle the necessary temperature has been attained
throughout the load and this is held for a minimum time.
NOTE: Calibration, which should not be confused with validation, compares the temperature set on the
dial to the temperature in the chamber. This is normally checked when the autoclave is serviced.