INDUSTRIAL OPERATION AND VALIDATION OF A DOUBLE DOOR AUTOCLAVE
PRESENTED BYB.KARTHIKM. Sc MICROBIOLOGY
INTRODUCTION OF AN AUTOCLAVE:- An autoclave is a pressure chamber used to carryout industrial processes requiring elevated temperature and pressure different from ambient air pressure. It is used for the sterilization of media, glass were, gloves, gowns etc.,It is a moist type of sterilization unit.Normal temperature and pressure required for autoclave is 121C and 15lbs pressure.
Aim:- To operate and validate the industrial double door autoclave.(Pharmalab)
Objective:-Operate and Validate the industrial double door auto clave. Dehydrated media, glass were and accessories sterilization for analysis .
HOW DOES AN AUTOCLAVE WORKS Steam enters the chamber jacket, passes through an operating valve and enters the rear of the chamber behind a baffle plate. It flows forward and down through the chamber and the load, exiting at the front bottom. A pressure regulator maintains jacket and chamber pressure at a minimum of 15 lbs, the pressure required for steam to reach 121C (250F). Overpressure protection is provided by a safety valve.
What is validation ? Validation shows that a process (or item of equipment) does what it is claimed to do.Why validate? Need to show that sterile equipment is being taken into sterile core - do not want to contaminate production (which could lead to adulterated product)For micro testing (both bacteriological and Viable Environmental Monitoring), using sterile equipment helps reduce false positives.Need to show that decontamination cycles are effective
How To Validate The usual URS. IQ, OQ, PQ.This discussion will focus on PQ.IQ: does equipment meet the URS requirements? Is everything that was on the box, in the box? Is the unit installed properly. Are support programs in place for ongoing operation of A/C?OQ: does the A/C operate properly? Does the unit hold temp and pressure correctly?PQ: validation of autoclave cycles and loading patterns need to show sterilization.
What is D value?Refers to decimal reduction time - The time required at a certain temperature to kill 90% of the organisms being studied. Thus after an organism is reduced by 1 D, only 10% of the original organisms remain. Dependant on microbe and initial numbers. E.g. D value of 1.5 = 1CFU .What is Z value?Refers to the temperature change required to produce a 1 log reduction in D value.What is F0?The number of minutes to kill a specified number of microbes with a Z value of 10oC at a temp of 121.5oC.Often confused with the time the chamber is held at elevated temperature and pressure and in practice is the same thing.
OverkillUse many more microbes than would find on items typically autoclaved. Negates the need to test sample for bio load before running the cycle.Use a sterilisation time exceeding what is necessary to kill a large number of microbes. Negates the need to determine D value of microbe.
Procedure to operate an autoclave :- Switch on auto clave Fill the water tank with water line Switch On the heater switch Open the door 1 Load the chamber with media or glass were or with accessories Arrange the sensors as we required Close the door- 2 login with user id and password
Select the type of recipe Start the cycle Set up the strip chartObserve the temperatures on the displayAfter completion of cycle open the door 2Give print to the printer Calculate the F0- value, D-value and Z- value with the use of computer software
Recipes for validation of a double door autoclaveThere are mainly 5 recipes present in the validation of a double door auto clave. They are :-VLT (Vacuum leak test)Bowie dick test cycleSteam In Process (SIP)Gravity test cycleHPHV test cycle
Vacuum leak test:-Vacuum leak test is performed to check any leakage in autoclaves. This test is also called chamber integrity test. In VLT, first vacuum is applied inside the chamber and then wait for the vacuum to reach at set vacuum point then hold the vacuum for required time and when hold time completed vacuum breakage occurs. After that check the results whether autoclave leakage is within the limits or not.
PARAMETERS:-Chamber level test vacuum :- -0.007barStabilization time :- 5minVacuum hold time :- 10minAcceptable leakage:- 0.0013bar/minPrint interval :- 60secFrequently test :- 15 days 2 daysAcceptable criteria :- 0.0013 bar/MinNo. of cycles run for validation :- 3
Bowie Dick test :-This is the cycle done for the removal of contaminated air pockets in an autoclave.In this the chemical indicator Bowie Dick chemical indicator papers will be used.The Bowie Dick indicator paper will turn from yellow to black(or) brown in colour.
Bowie Dick test papers
Parameters :-Pre vacuum :- -0.700Bar Pre vacuum hold time :- 01 minPre pressure :- 0.300BarPre pressure hold time :- 01 minHeat up 1 :-107CHeat up 1 hold :- 01minHeat up 2 :- 112Heat up 2 hold :- 01minHeat up 3 :- 115Heat up 3 hold :- 01 minHeat up 4:-117Heat up 4 hold :- 1minSterile hold temp :- 121.3
Control band :- 000.3Sterile hold time :- 11 minSterile hold reset :- 120.6 Sterile hold stop count :- 121Post vacuum :- -0.700BarPost vacuum hold time :- 02 minDoor open temp :- 90Print interval :- 60 secControl temp :- min for all
SIP (Steam In Process) cycle:-It is the cycle done for vent filter sterilization of an autoclave.It is the test done once per a monthAcceptance criteria :-For SIP of filter temperature should be more than 121C during sterilization.
Parameters :-Heat up 1 :-107CHeat up 1 hold :- 01minHeat up 2 :- 112CHeat up 2 hold :- 01minHeat up 3 :- 115CHeat up 3 hold :- 01 minHeat up 4:-117CHeat up 4 hold :- 1minSterile hold temp :- 122CSterile hold time :- 15 min
Sterile hold reset :- 120.6C Sterile hold stop count :- 121CSterile temp over shoot :- 130 CPost vacuum :- -0.700BarPost vacuum hold time :- 02 minDoor open temp :- 90Print interval :- 60 sec
Gravity cycle :- This is the cycle used for the sterilization of the media and water.This test is done as per our requirement.Temperature should be in between 121 124 during sterilization hold time.The steam pressure should be 1.04-1.24 bar during the sterilization hold time.In this the chemical indicator paper and biological indicator will be used.The biological indicator is Geobacillus sterothermophillus.This is a spore producing bacteria which can resist up to 120C temperature.
Parameters :-Heat up 1 :-105CHeat up 1 hold :- 02minHeat up 2 :- 108Heat up 2 hold :- 02minHeat up 3 :- 112Heat up 3 hold :- 02 minHeat up 4:-117Heat up 4 hold :- 03minSterile hold temp :- 121.3Control band :- 000.3Sterile hold time :- 20 minSterile hold reset :- 120.5
Sterile hold stop count :- 121Sterile temp over shoot :- 124Jacket control temp :- 121Door open temp :- 96Print interval :- 60 secControl temp :- min for all
Biological indicator for gravity cycle Geobacillus sterothermophillus Before testAfter test
HPHV cycle :-High pressure and high vacuum test cycle is used for the sterilization of a glass were, accessories etc.,Temperature should be in between 121 124 during sterilization hold time.The steam pressure should be 1.04-1.24 bar during the sterilization hold time.
Parameters :-Pre vacuum :- -0.700Bar Pre vacuum hold time :- 02 minPre pressure :- 0.300BarPre pressure hold time :- 02 minHeat up 1 :-107CHeat up 1 hold :- 02minHeat up 2 :- 112CHeat up 2 hold :- 02minHeat up 3 :- 115CHeat up 3 hold :- 02 minHeat up 4:-117CHeat up 4 hold :- 1minSterile hold temp :- 121.3CControl band :- 000.3
Sterile hold time :- 30 minSterile hold reset :- 120.6C Sterile hold stop count :- 121CSterile temperature over shoot :- 124CPost vacuum :- -0.700BarPost vacuum hold time :- 02 minDoor open temp :- 90Print interval :- 60 sec
Observations and Results:Vacuum leak test:
Run - IRun - IIRun IIIDate25/05/201625/05/201625/05/2016Cycle start time14;53;5015:16:3715:59:19Vacuum hold start time15:01:1615:24:0416:06:49Vacuum hold end time15:11:1615:34:0416:16:49Cycle completion time15:12:3515:35:2316:18;09Vacuum hold start pressure-0.723 bar-0.724 bar-0.725 barVacuum hold end pressure-0.717-0.717-0.717Actual leakage0.006bar0.007bar0.008bar
Bowie Dick test Run IRun IIRun IIIDate25/05/201625/05/201625/05/2016Cycle start time16:38:0118:04:2019:11:44Sterile hold start time17:18:5818:29:2519:36:13Sterile hold end time17:29:5818:40:2519:47:13Cycle completion time17:47:3219:00:0820:05:42
No leaks observed Bowie Dick kit turns blue colour to black colour
Steam in process (SIP) test:
Run IRun IIRun - IIIDate27/05/201627/05/201627/05/2016Cycle start time 13:36:0014:29:4216:14:40Steril