Sterilization can be accomplished by either physical or chemical means. The principal physical means is autoclaving, using a Steam sterilizer; other physical methods include boiling and dry heat. Chemicals used for sterilization include the gases ethylene oxide and formaldehyde, and liquids such as glutaraldehyde.F
Of all these forms of sterilization, steam sterilization by way of autoclaving is the fastest, most reliable and hence; most commonly used within the FDA, MHRA, WHO and EU zones of influence. It must always be remembered that it is also one of the easiest processes to get wrong. This is why regulators will nearly always scrutinize and ask about validation methods used in autoclave validation and qualification activities.
Autoclave validation / Qualification must follow the routine validation / Qualification document string of;
Autoclaving is the most effective and most efficient means of sterilization. All autoclaves must go through the GMP process of autoclave validation / Qualification during which, the various programs are verified as conforming to the requirements detailed in the User Requirement Specification (URS). They operate on a time/temperature relationship. These two variables are extremely important. Higher temperatures ensure more rapid killing. Some standard temperature/pressures employed are 115ºC/10 p.s.i., 121ºC/15 p.s.i., and 132ºC/27 p.s.i. Longer times are needed for larger loads, large volumes of liquid, and more dense materials. Autoclaving is ideal for sterilizing biohazardous waste, surgical dressings, glassware, many types of microbiologic media, liquids, and many other things. When proper conditions and time are employed, no living organisms will survive a trip through an autoclave.
The thermal resistance of specific microorganisms is characterized by “D”–values and “Z”–values. A D-value is the time in minutes, at a specific temperature, to reduce the surviving microbial population by 1 – log. A Z-value is the temperature change required to result in a 1-log reduction in D-value. Other time measurement variables pertaining to thermal resistance are F-values and Fo-values. An Fo-value is the number of minutes to kill a specified number of microorganisms with a specified Z-value at a specific temperature. An Fo– value is the number of minutes to kill a specified number of microorganisms with a Z-value of 10° C (50° F) at a temperature of 121.1° (250°F).
It is not unusual to find people thinking 121° C is the temperature for sterilization. In the early days of autoclave validation, steam for sterilization at standard temperature was used in order that studies could be accurately compared, the temperature chosen was a nice round figure of 250deg F (121.1° C). The Fo-value can be determined as per the following
Fo = 10 (T – 121.1)/10
Where T = temperature (° C) and Fo = equivalent sterilization time (minutes)
So given a Bioburden of 1215 CFU, with a D-value of 1.6 min/log at 121.1°C and a required SAL of 10-6.
Then: Log (1215) = 3.08
Loge reduction = 3.08 log + 6 log = 9.08 log.
Ideal Cycle at 121.1°C (250°F) = (9.08 log)(1.6 min/log) = 14.53 minutes.
Moist heat sterilization (or autoclaving) is conducted by supplying dry, saturated steam under pressure to an autoclave. The energy (heat) from the condensation of steam on the items in the sterilizer will kill the present microorganisms by irreversible damage of cell components.
The effectiveness of steam sterilization in the form of moist heat sterilization process increases considerably when air is removed before adding steam to the chamber. Obtaining a vacuum can be difficult, resulting in limited capability of the steam to penetrate into cavities of instruments etc. The use of biological indicators during autoclave validation / Qualification is therefore recommended for monitoring allowing the conditions at different points in the sterilized goods to be assessed.
Biological indicators, used during autoclave validation include preparations of selected microorganisms (bacterial spores) with high resistance towards specific sterilization methods. The bacterial spores are deposited on a carrier, e.g. filter paper, which is wrapped in a suitable package making the system ready for use and with defined resistance characteristics. The inactivation of the biological indicator indicates an effective sterilization process. Whether inactivation has been obtained is determined by cultivation after exposure.
To ensure and to test that an acceptable amount of moisture is present in the steam supply for autoclave validation. For little amount of moisture there is a chance of superheating may occur. Even too little moisture may prevent sterilizing conditions in the chamber. Steam with a dryness fraction of 0.99 consists of 99% steam and 1% water. Similarly, steam with a dryness fraction of 0.95 consists of 95% steam and 5% water. The dryness value of the steam should be equal to or greater than 0.9 for porous loads or 0.95 where metal loads are processed.
There are quite a few reasons why superheated steam is not as suitable for use in steam autoclave validation and general use. In general applications, steam with a large degree of superheat is of little use because it:
a) Gives up little heat until it has cooled to saturation temperature.
b) Creates temperature gradients as it cools to sat temperature.
c) Lower rates of heat transfer whilst the steam is superheated.
d) Requires larger heat transfer areas.
Positioning of the thermocouples (t/c's) during autoclave validation / Qualification or indeed in any GMP temperature mapping exercise is all about appreciating what is adding or subtracting heat from the room or cabinet being qualified.
In the case of temperature mapping during autoclave validation / Qualification, heat is added in the form of pressurized wet steam, anything that can affect the distribution of the incoming steam, can affect uniformity of temperature. Conversely anything that can take heat away from the chamber can affect temperature uniformity.
Let me say at this stage if you want to be pedantic and put t/c’s down the drain, the mapping exercise will probable fail. However you are there to verify that product will be sterilized, and product is never placed down the drain. Only the designated product containment area has to be verified.
If this is new installation, then get hold of the Factory Acceptance Test (FAT). In the FAT the chamber is subjected to detailed temperature transfer studies.
Even distribution of the in-coming steam can be verified by placing a thermocouple sensor (t/c) in each of the eight corners in the autoclave and one in the cabinet centre. (9 t/c’s)
Cooling due to heat loss will be maximum the further away you are from the steam inlet and the closer you are to metal that will conduct heat out of the chamber. That is usually, the door, or doors if double sided. The drain is also a heat sink that conducts heat out of the chamber. One t/c should be placed as close to the drain as product would be, when the autoclave validation is in normal use and another placed alongside the cabinet product temperature probe. This gives us an additional 2 t/c’s, bringing the total for a standard sized autoclave validation task to 11 t/c’s.
This is normally considered sufficient for 1.5 to 2.5 m3 autoclave validation tasks. Any bigger and I would concentrate on heat loses i.e. add t/c’s to the top and bottom of the doors and or end wall.
It is most important to understand that it is impossible for autoclave validation / Qualification to be successfully executed while using none validated steam.
Your steam must be validated for – dryness – none condensable gases.
Another GMP essential is to carry out pre and post mapping, calibration of your thermocouples. These should be calibrated against test standard instruments whose calibration is traceable to national standards, and for which you have valid current calibration certification.
Risk Mitigation in Validation template (Issue 11) $125.00 The Risk and Part 11 Validation Risk Assessment(VRA) protocol is becoming the most important document in the validation online train. The VRA reassures the regulators that you have looked at specific equipment functionality and considered the appropriate level of validation that is required. You have also considered various aspects of its use and the implications of any malfunctions. From the results of this exercise the scope of all validation activity can and must be justified. This is a robust and simple to execute document, one that will lead you through the process and deliver a result that can be used as the foundation for your Risk Mitigation in Validation template.
This VRA now includes the assessment table for categorizing and documenting the new 21 CFR Part 11 guidance ruling on what predicate data must be stored in a Part compliant system, along with the new broadsheet to establish your new database of part 11 records. (now mandatory).
This Pharmaceutical Validation combination protocol has been produced in response to several hundred reader suggestions we received in our ‘Suggestions Section’. It has been carefully designed to make it the preferred choice for Process and Laboratory stand alone equipment and associated standard operating procedure TEMPLATE. The associated medical device validation Master Plan template is interactive, easy to use and suitable for all mixes of equipment with and without software. Where temperature mapping is required it will included in this section. This of course may well be a Risk Mitigation in Validation risk Assessment template. As per 21 CFR Part 11.
The Installation Qualification Template or IQ section establishes documented verification that all medical device validation template online executables are catered for and that key aspects of the equipment Qualification adhere to approved design Qualification template and the recommendations of the manufacturer have been suitably considered. The Operational Qualification Template section establishes that there is documented verification that the installed system functions as specified and that there is sufficient documented process validation A executables to demonstrate this. The PQ section develops documented evidence that all the requirements specified in your corporate validation 4U manual have been verified as operating consistently and exactly as specified in the medical device validation template; The User Requirements Specification template
The Standard Operating Procedure template (SOP) used to generate this installation Qualification template (IQ), takes you through the process line by line, chapter by chapter. It really is unique to find a SOP document so easy to use, all the work is done for you. All the documents are detailed, all the drawings listed and all the checks and tests detailed. The final product is a professional and comprehensive Installation Qualification template. One that you can produce in less than 60 minutes. Yes, think about it, we all know how long producing IQ documents has taken in the past, whether for current Good Manufacturing Practice use medical device use or a simple installation qualification template.
You will find this validation online intuitive Operational Qualification template (OQ) delightfully simple and straightforward to use, as it takes you through the process of customization of your Operational Qualification Protocol template. Following the attached SOP will quickly and smoothly convert your template into an equipment specific 21 CFR Part 11 compliant Installation Qualification Template. The OQ template comes complete with all the standard test scripts, more specialist test scripts can be found listed below. These can easily be pasted into the standard OQ, allowing you to quickly build your own fully detailed and referenced company bespoke Operational Qualification Protocol, along with the installation qualification template.
The SOP used to generate this IQ, takes you through the process line by line, chapter by chapter. It really is unique to find a SOP document so easy to use, all the work is done for you. All the documents are detailed, all the drawings listed and all the checks and tests detailed. The final product is a professional easy to use and comprehensive autoclave validation template validation Installation Qualification Protocol. One that you can produce in less than 60 minutes. Yes, think about it, we all know how long producing IQ documents has taken in the past.
This autoclave Validation template Operational Qualification SOP and Protocol, can be simply and quickly converted (using find replace techniques) into your own document. Following the attached SOP can further convert it into your completely simply to use bespoke, clever & intuitive Operational Qualification Protocol, speedily raised and ready to issue for approval signatures, prior to execution. This protocol includes all test scrips and temperature mapping as part of the OQ.
The SOP used to generate this PQ, takes you through the process line by line, chapter by chapter. It really is unique to find a SOP document so easy to use, all the work is done for you. All the documents are detailed, all the drawings listed and all the checks and tests detailed. Completion of this Autoclave Validation Template Validation performance validation protocol allows you to proceed to the process validation stage.
This package of documents is ideal for those who are in the procurement stage for one or more new or refurbished autoclaves. Follow the Validation plan (VP) and use it to ensure all eventualities in the procurement process are identified and planned for. Ensure resources are allocated for all minor and major support tasks. Use the DQ to verify that this design of autoclave is capable of fulfilling all your functional, GMP and safety requirements. If all your autoclaves are the same then one VP, DQ and URS may be all that is required. Then it is quite simple to copy over individual IQ, OQ and PQ protocols from the document package; which on execution will verify that all the requirements as documented in the URS have been fully satisfied.
This autoclave validation template package contains:
VMP, VP, URS, VRA, DQ, IQ, OQ & PQ.