1. EQUIPMENT VALIDATION - Autoclave Dr. L. Prabakaran Prof. & Head R. R. College of Pharmacy Bangalore -90.

2. History of Validation 1970’s- The term validation started gaining attention 1970’s- The term validation started gaining attention 1980’s –Process validation is here to stay- FDA Books on Process validation started appearing 1980’s –Process validation is here to stay- FDA Books on Process validation started appearing 1990’s – Emphasis on Validation 1990’s – Emphasis on Validation Current – As a GMP concept Current – As a GMP concept

3. Introduction Next to GMP, Validation is most used term today and has become a catchy word. Next to GMP, Validation is most used term today and has become a catchy word. Validation is no more a high sounding word. It has become reality and part and parcel of GMP. Validation is no more a high sounding word. It has become reality and part and parcel of GMP. Validation makes the life livelier. Validation makes the life livelier. Today the term is defined in the law. Today the term is defined in the law. New Schedule M deals with it. New Schedule M deals with it.

4. Validation is one which has been proved to do what it purports to do. Validation is one which has been proved to do what it purports to do. The proof of Validation is obtained through collection and evaluation of the data preferably beginning from the process. The proof of Validation is obtained through collection and evaluation of the data preferably beginning from the process. Validation is documented common sense. Validation is documented common sense.

5. Reasons for Validation Quality Assurance Quality Assurance Economics Economics Reduction in the cost associated with process monitoring sampling and testing Reduction in the cost associated with process monitoring sampling and testing Qualify the goals towards Parametric release Decrease or eliminate product rejections Qualify the goals towards Parametric release Decrease or eliminate product rejections Compliance - FDA’s requirement Compliance - FDA’s requirement

6. Means a written plan stating how validation will be conducted, including test parameters, product characteristics, production equipment, and decision points on what constitutes acceptable test results. Validation Protocol

7. Autoclave Autoclaves use pressurized steam to destroy microorganisms, and are the most dependable systems available for the decontamination of laboratory waste and the sterilization of laboratory glassware, media, and reagents. Autoclaves use pressurized steam to destroy microorganisms, and are the most dependable systems available for the decontamination of laboratory waste and the sterilization of laboratory glassware, media, and reagents. Autoclaves should be tested periodically with biological indicators like cultures of Bacillus stearothermophilus to ensure proper function. Autoclaves should be tested periodically with biological indicators like cultures of Bacillus stearothermophilus to ensure proper function. This method of sterilization works well for many metal and glass items but is not acceptable for rubber, plastics, and equipment that would be damaged by high temperatures This method of sterilization works well for many metal and glass items but is not acceptable for rubber, plastics, and equipment that would be damaged by high temperatures

8. Autoclave

9. Autoclaves, or steam sterilizers essentially consist of following: i) A cylindrical or rectangular chamber, with capacities ranging from 400 to 800 liters. ii) Water heating system or steam generating system iii) Steam outlet and inlet valves iv) Single or double doors with locking mechanism. v) Thermometer or temperature gauge vi) Pressure gauges

10. For porous loads (dressings) sterilizers are generally operated at a minimum temperature of 134 0 C, and for bottled fluid, sterilizers employing a minimum temperature of 121 0 C are used. For porous loads (dressings) sterilizers are generally operated at a minimum temperature of 134 0 C, and for bottled fluid, sterilizers employing a minimum temperature of 121 0 C are used. Ensure that there should be sufficient water in the autoclave to produce the steam. Ensure that there should be sufficient water in the autoclave to produce the steam. The stages of operation of autoclaves include air removal, steam admission and sterilization cycle (includes heating up, holding/exposure, and cooling stages). The stages of operation of autoclaves include air removal, steam admission and sterilization cycle (includes heating up, holding/exposure, and cooling stages). Operation

11. AUTOCLAVE VALIDATION – WHAT IS REALLY REQUIRED?

12. Load Types Autoclaves are used to sterilize several different types of loads: Solid – metal, glass, plastic Solid – metal, glass, plastic Porous – linen, gowns, paper, gauze, complex instruments, hollow tubes Porous – linen, gowns, paper, gauze, complex instruments, hollow tubes Liquid – water, saline, media Liquid – water, saline, media Laboratory waste – Petri dishes, sample bottles, syringes Laboratory waste – Petri dishes, sample bottles, syringes

13. System supplies Most autoclaves are electrically powered and all steam autoclaves require some form of a water supply to produce steam. Most autoclaves are electrically powered and all steam autoclaves require some form of a water supply to produce steam. The electrical supply must be able to handle the maximum peak power of the autoclave. The electrical supply must be able to handle the maximum peak power of the autoclave. The drainage system must be able to handle the maximum flow of water and condensate and should be able to tolerate 100 °C. The drainage system must be able to handle the maximum flow of water and condensate and should be able to tolerate 100 °C.

14. Commissioning Commissioning tests should include checking: that the electrical supply is suitable for the autoclave that the electrical supply is suitable for the autoclave that the temperature and humidity are sufficiently well controlled that the temperature and humidity are sufficiently well controlled that there is no interference to or from other equipment that there is no interference to or from other equipment the accuracy of indicators has been certified to national standards the accuracy of indicators has been certified to national standards

15. the temperature and pressure of the chamber during a sterilization cycle are within specifications the temperature and pressure of the chamber during a sterilization cycle are within specifications the leakage into the chamber during a vacuum cycle does not exceed the specified maximum the leakage into the chamber during a vacuum cycle does not exceed the specified maximum that there is no leakage of steam, water or effluent at any point during the sterilization cycle that there is no leakage of steam, water or effluent at any point during the sterilization cycle

16. Routine Checking Routine checking may be broken down into daily, weekly, quarterly and annual checks. Routine checking may be broken down into daily, weekly, quarterly and annual checks. Daily checks should be performed by the operator and should include the observation of all indicators for “normal” values. Daily checks should be performed by the operator and should include the observation of all indicators for “normal” values. On a weekly basis safety checks should be performed such as inspection for leaks and electrical wiring faults, inspection of door seals, correct of operation of door interlocks and high temperature and low water alarms. On a weekly basis safety checks should be performed such as inspection for leaks and electrical wiring faults, inspection of door seals, correct of operation of door interlocks and high temperature and low water alarms.

17. Routine checking may be broken down into daily, weekly, quarterly and annual checks. Routine checking may be broken down into daily, weekly, quarterly and annual checks. Daily checks should be performed by the operator and should include the observation of all indicators for “normal” values. Daily checks should be performed by the operator and should include the observation of all indicators for “normal” values. On a weekly basis safety checks should be performed such as inspection for leaks and electrical wiring faults, inspection of door seals, correct of operation of door interlocks and high temperature and low water alarms. On a weekly basis safety checks should be performed such as inspection for leaks and electrical wiring faults, inspection of door seals, correct of operation of door interlocks and high temperature and low water alarms.

18. Documentation The results of all commissioning tests and performance qualification tests should be fully recorded, reviewed, accepted and archived by the relevant personnel in the organization that uses the autoclave. The results of all commissioning tests and performance qualification tests should be fully recorded, reviewed, accepted and archived by the relevant personnel in the organization that uses the autoclave.

19. Heat distribution studies: The intent of this study is to demonstrate the temperature uniformity & stability of the sterilizing of the sterilizing medium throughout the sterilizer. The intent of this study is to demonstrate the temperature uniformity & stability of the sterilizing of the sterilizing medium throughout the sterilizer. Temperature distribution studies should be conducted on both the empty and loaded chambers, with maximum & minimum load configurations. Temperature distribution studies should be conducted on both the empty and loaded chambers, with maximum & minimum load configurations. A narrow temperature uniformity range is required, generally variation of less than ±1ºC of the mean chamber temperature. A narrow temperature uniformity range is required, generally variation of less than ±1ºC of the mean chamber temperature. If equipment is to be used in the terminal sterilization of products uniformity of ±0.5ºC of the mean is desirable. If equipment is to be used in the terminal sterilization of products uniformity of ±0.5ºC of the mean is desirable.

20. Object mapping; The study includes determination of slowest to heat zone within an object such as filter housings, hoses, containers filled with the stoppers. The study includes determination of slowest to heat zone within an object such as filter housings, hoses, containers filled with the stoppers. Heat penetration studies; the intent of the study is to determine the coolest points within a specified load & configuration. the intent of the study is to determine the coolest points within a specified load & configuration.

21. Microbiological challenge studies: Biological castles are employed during heat penetration situations in order to demonstrate the degree of process lethality provided by the sterilization cycle. Biological castles are employed during heat penetration situations in order to demonstrate the degree of process lethality provided by the sterilization cycle. Calibrated biological indicators utilized are bacillus stearothermophilus & clostridium sporogenes, spore-forming bacteria because of their relatively high heat resistance. Calibrated biological indicators utilized are bacillus stearothermophilus & clostridium sporogenes, spore-forming bacteria because of their relatively high heat resistance.

22. MONITORING STERILIZATION PROCEDURES Sterilization procedures can be monitored routinely using a combination of biological, chemical and mechanical indicators as parameters. Sterilization procedures can be monitored routinely using a combination of biological, chemical and mechanical indicators as parameters. Biological Indicators Monitoring the sterilization process with reliable biological indicators at regular intervals is strongly recommended. Monitoring the sterilization process with reliable biological indicators at regular intervals is strongly recommended. Measurements should be performed with a biological indicator that employs spores of established resistance in a known population. Measurements should be performed with a biological indicator that employs spores of established resistance in a known population.

23. Chemical Indicators Chemical indicators include indicator tape or labels, which monitor time, temperature and pressure for steam sterilization, and time and temperature for dry- heat sterilization. Chemical indicators include indicator tape or labels, which monitor time, temperature and pressure for steam sterilization, and time and temperature for dry- heat sterilization. These indicators should be used on the inside and outside of each package or container. These indicators should be used on the inside and outside of each package or container. External indicators are used to verify that items have been exposed to the correct conditions of the sterilization process and that the specific pack has been sterilized. External indicators are used to verify that items have been exposed to the correct conditions of the sterilization process and that the specific pack has been sterilized. Internal indicators are placed inside a pack or container in the area most difficult for the sterilization agent to reach. This is the indicator that tells if the item has been sterilized. Internal indicators are placed inside a pack or container in the area most difficult for the sterilization agent to reach. This is the indicator that tells if the item has been sterilized.

24. Dry Heat Sterilization IndicatorsPrincipleDevice Parameter monitored Chemical Temperature sensitive colored solution Browne’s tube Temperature, time Temperature sensitive chemical A temperature sensitive white wax concealing a black marked Temperature Biological Temperature sensitive microbes Bacillus subtilis D value

25. Moist Heat Sterilization IndicatorsPrincipleDevice Parameter monitored Chemical Temperature sensitive colored solution Browne’s tube Temperature, time Steam sensitive chemical A device which is impregnated into a carrier material Saturated steam Biological Temperature sensitive microbes Bacillus Stearotherophilus D value