AUTOCLAVE VALIDATION PROTOCOL

Validation of the Autoclave is classified into the following
1.0 OQ – Operational Qualification
2.0 PQ – Performance Qualification
The validation is being taken up to cater to the new requirements of the GMP. Since it is already in use only OQ and PQ will be considered.
VALIDATION TEAM:

OPERATIONAL QUALIFICATION PROTOCOL (OQ)
1.0 PURPOSE :
To demonstrate and document that the operations
of the Autoclave take place as specified .
2.0 SCOPE :
Autoclave xxxxx will be qualified to meet OQ.
3.0 RESPONSIBILITY :
Microbiologist, Manager Q.C
4.0 PROCEDURE :
This should be performed by external agency like IIME.
4.1 Verify the following as per instrument operating procedure and calibration certificate kept in place before validation.
4.1.1 : Temperature display of Autoclave.
4.1.2 : Compound pressure gauge of Autoclave.
Acceptance criteria :
All calibration data found to be within the acceptable norms of calibration certificate.
4.2 Calibration of Thermocouples :
Calibrate all the thermocouples of data logger before and after the validation using standard thermometer and also made available party’s calibration certificates.
Acceptance criteria:
The variation between the temperature of thermocouples and the standard thermometer found to be within the acceptance criteria.
4.3 Heat Distribution Studies
Carry out heat distribution studies by using a multi-point data logger and maintain holding time for 15 minutes at 15 lbs. by fixing all the 12 probes as per diagram-1. Record the temperature and lag time of each probe as per Annexure –1 & 2.
Acceptance criteria
All probes must reach temperature 121-124°C and pressure must be within 15 to 18 lbs for 15min cycle.
Diagram-1

Probe No.1 to 12 inside the chamber

Load Pattern
Maximum Load
Load with all the glassware and media filled upto 70%, of the chamber and the details are as follows.
Test-1 : 250ml Conical flasks = 12 Nos with media, 13 Nos without media, 500ml Conical flasks with media = 4nos, 1000ml Conical flasks with media =4nos, Pipette10ml=10nos,Pipette 2ml=10nos, Pipette 5ml=10nos, Pipette 1ml = 10nos,100ml
bottles=20nos ,Filtering unit=10nos, Test tubes =25nos
Minimum Load
Load with all the glassware and media required for a day’s analysis (average).
250ml Conical flasks with media = 6nos, 500ml Conical flask with media – 3 nos., 1000ml Conical flask - 1 Pipette 10ml= 10nos, Pipette 2ml= 10nos, Pipette 5ml= 10nos, Pipette 1ml=10nos, Bottles=10nos, Test tubes - 25nos.
Record the temperature and lag time and tabulate as per annexure 1.

PERFORMANCE QUALIFICATION PROTOCOL (PQ)
1.0 PURPOSE :
To provide a performance qualification protocol for Autoclave.
2.0 SCOPE :
Specified to Autoclave xxxxx.
3.0 RESPONSIBILITY :
Microbiologist, Manager Q.C
4.0 PROCEDURE :
4.1 Heat Penetration Studies
Carry out the heat penetration studies by using a multi point data logger for the following loads mentioned. Record the temperature and lag time if any as per annexure 1 and 2.
Acceptance criteria:
All 12 probes must reach temperature 121°C to 124°C and pressure must be within 15 to 18 lbs. for 15 min cycle.
4.2 Microbial limit test :
Incubate the sterilised media flask or tubes from any one Maximum and Minimum load of heat penetration studies and observe for nutritive properties. Bacteria: 30 – 35 °C for 72 hrs, Fungi : 20 – 25°C for 120 hrs
Acceptance criteria:
No microbial growth should be observed i.e. Negative control and Nutritive properties of media must pass
4.3 Microbial challenge test :
Keep ampoules containing spores suspension of Bacillus stearothermophilus 106 ­ population at various location of the autoclave along with probes and maintain the sterilisation temperature at 15psi and 121°C during the heat penetration studies, once on the maximum load.
Acceptance criteria:
Autoclaved ampoules containing Bacillus stearothermophilus spores suspension ampoules should not show any colour change after five days of incubation.
5.0 DOCUMENTATION :
5.1
Master Instrument used for validation of autoclave
Institutes name and address carrying out calibration.
Standard calibrating instrument name and number.
Instrument certified against (Instrument of national or international standards)
Date of calibration and validity period of calibration.
Training certificate of persons (External agency) carrying out validation.
6.2 Autoclave being calibrated :
All temperature readings for autoclave being validated should be collected from the approved external agency like IIME.
Validation report with observed any error, statement of calibration and next validation due.
7.0 FREQUENCY :
Once in a year until and unless no change in autoclave. In case of any change, the autoclave must be revalidated

8.0
CONCLUSION : Finally conclusion should be drawn based on the results of above tests and documented.

HVAC QUALIFICATION.

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HVAC QQUALIFICATION INSTALLATION.

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HVAC OPERATIONAL QUALIFICATION.

This easy to use HVAC Operational Qualification SOP and Protocol, can be simply and quickly converted (using find replace techniques) into your own company bespoke document. Now you can purchase these document in a suite DQ to PQ, getting you ready at express speed to qualify a system. It has never been quicker or simpler.

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HVAC PERFORMANCE QUALIFICATION.

This fully formatted HVAC Performance Qualification is the last in the chain of qualifying tests that HVAC systems are subjected to, before being considered qualified. However environmental qualification is required to verify air quality.

STEAM QUALITY TESTING.

Steam Quality.
A continuous supply of saturated steam is required for steam sterilization and for humidification in certain EO sterilizers. Too high a level of non condensible gases will prevent the attainment of sterilization; too little moisture carried in suspension may allow the steam to become superheated during expansion into the chamber, while excess moisture may cause damp loads. Where steam systems are either routinely or irregularly shut down, large quantities of air will be present in the distribution system on restarting. It is recommended that in such circumstances a comprehensive and validated venting procedure should be applied and testing for steam quality is appropriate.

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Non-condensible gases

Non-condensible gases result from the water from which the steam is generated. These gases will usually be air, though carbon dioxide may be present, caused by certain water treatment processes, typically water softeners. This is exacerbated by excessive aeration that can exist in many pharmaceutical water treatment plants where water is constantly recirculated and sprayed into the top of storage vessels. The effect of such gases being present in the steam supply to a sterilizer can be the same as air, none sterilization of the volume they occupy.

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Superheated steam.

Superheated steam is steam at a temperature above its boiling point for its pressure. Superheated steam is a clear colour-less gas that will not condense until its temperature drops to its boiling point. Until this occurs the moisture necessary for sterilization cannot be produced and therefore presents a risk to the process. Superheated steam acts as hot air and requires sustained high temperatures and long hold times before sterilization can occur.

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Dryness Value Test.

Wet steam is undesirable as it has less energy than dry steam and more importantly can cause wet loads. The packaging used for sterile products prevents reinfection when dry, but its bacterial retentive properties will be adversely affected by the presence of moisture. Wet loads can be considered to be un-sterile. The dryness fraction describes how dry steam is, with a value of 1 representing steam that is 100% dry, and therefore free of entrained moisture. Steam with a dryness fraction of 0.99 consists of 99% steam and 1% water. If we measure the latent heat present in steam that has a dryness fraction of 0.99 we will find that it possesses 99% of the full quotient of latent heat.

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VALIDATION RISK ASSESSMENT ASSESSMENT RATIONALE

The question - how much validation - has been an open subject for many years. Numerous companies brought in validation consultants, and or regulatory compliance experts, to guide them through these problems, but every so often, there would be clashes, or at least variations in opinion, in what the appropriate scope and depth of validation was. It was accepted by the regulators that there was going to be equipment used, that full validation was inappropriate for, yet the design or misuse of that piece of equipment, could be critical to product quality. This cannot be acceptable, so how do we prevent it occurring?

When we move away from these simple devices, to the average type of production equipment, here we find a range of equipment varying from purely mechanical (but intricate) equipment, to mechanical equipment assisted and or monitored and or managed by some degree of electronics. Then at the very pinnacle of validation we have the mainly electronic systems running extremely complex sophisticated software programs. All these systems play a major role in the product manufacturing process, they all require thorough validation using the existing framework of IQ – OQ - PQ, but they do not all require the same level of validation. How do we define the appropriate level?

First we have to look at validation and how the level of validation can be varied independently of the scope. If we list all the tasks we consider essential to complex software driven equipment validation, then drop off the tasks we consider inappropriate to the next level of equipment complexity, and so on. We appear to have four levels, with a significant step change between each of them.

• Mechanical/electronic/electrical/full life cycle software, validation.
• Mechanical/electronic/electrical/software, validation.
• Mechanical/electronic/electrical/software, calibration.
• None

The requirement for 21 CFR Part 11 compliance, can occur at all three of the upper levels of validation and so is addressed in the Validation Risk Assessment protocol as a separate requirement. To find that a piece of laboratory equipment requires the same degree of validation as a Distributive Control System may be disquieting, however any equipment that uses software alone, to automatically derive whether the product passes, or fails to pass, an inspection stage, must be subjected to FLCV. The laboratory equipment may have one such stage, the DCS will have many, however, one or many, the software lineage for each system, must follow similar methodologies.

Our Validation Risk Assessment (VRA) takes you through this assessment process and enables you to make a documented and justified decision as to the level of validation each piece of equipment will receive. This fulfils your obligation to ensure that all software is assessed, as to FLCV applicability.

Temperature Mapping Autoclave and Others

Positioning of the thermocouples in any temperature mapping exercise is down to judgment. A judgment that must be documented and justified.

In the case of an autoclave, 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.

Lets me say in this VALIDATION ONLINE BLOG, this stage if you want to be pedantic and put tc’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 in the drain. Only the designated product containment area has to be verified.

If this is new installation then get hold of the 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 (tc) in each corner of the autoclave (8 tc’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. If not double sided, then the end wall, and last but not least, the drain. The drain is a big source of heat loss. A tc should be placed as close to the drain, as product would be when the autoclave is in normal use. This gives us an additional 3 tc’s, bringing the total for a standard sized autoclave to 11 tc’s.

I have always considered this sufficient for 1.5 to 2.5 m3 autoclaves. Any bigger and I would concentrate on heat loses i.e. add tc’s to the top and bottom of the doors and or end wall.

It is most important to understand that it is impossible to validate an autoclave while using none validated steam.Your steam must be validated for – superheat – dryness – none condensable gases.

Make certain you do a pre and post mapping calibration, using instrumentation traceable to national standards.Happy mapping.