Autoclave Operation & Performance Testing

Safe autoclave use.

Environmental Health & Safety Author

06/26/2019 Added

880 Plays

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Information on safe operation and performance testing to ensure proper use and effective decontamination.

Demonstration of autoclave operation/performance testing.

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[00:00:01.024](energetic music)
[00:00:12.671]Autoclaves are used
[00:00:13.730]in many areas to sterilize materials.
[00:00:16.400]Due to the high heat and pressure created
[00:00:18.420]in autoclaves during operation, proper loading, use
[00:00:21.970]and unloading procedures must be followed
[00:00:24.300]to prevent burns and other accidents.
[00:00:26.690]Burns can result from physical contact
[00:00:28.930]with the structure of the autoclave,
[00:00:30.630]and steam burns can occur
[00:00:32.040]from contact with steam leaving the apparatus.
[00:00:35.330]Burns can also result from careless handling
[00:00:37.520]of vessels containing hot liquids.
[00:00:39.890]Explosive breakage of glass vessels
[00:00:41.890]during opening and unloading due to temperature stresses
[00:00:44.710]can lead to mechanical injury, cuts and burns.
[00:00:48.100]Autoclave performance for sterilization
[00:00:50.120]is dependent on proper use.
[00:00:52.610]This video provides guidance related to proper use
[00:00:55.530]and testing the performance of autoclaves
[00:00:57.580]with biological indicators.
[00:01:00.290]Gravity: in this mode, evacuation of air
[00:01:04.810]from the autoclave chamber
[00:01:06.180]prior to the sterilization portion of the run
[00:01:08.930]is accomplished by gravity air purge.
[00:01:11.520]A gravity cycle is appropriate for loads
[00:01:14.020]where air removal from porous materials
[00:01:16.280]or penetration of steam into wrapped
[00:01:18.740]or packaged items is not required.
[00:01:22.010]Vacuum cycle: in this mode, evacuation of air
[00:01:26.650]from the autoclave chamber
[00:01:27.990]prior to the sterilization portion of the run
[00:01:30.540]is accomplished by pulsing between pressure and vacuum.
[00:01:34.320]As the number of pulses, or prevacs increases,
[00:01:37.860]so does the efficiency of air removal
[00:01:40.050]and subsequent steam penetration.
[00:01:42.560]Vacuum mode is suitable for hard goods
[00:01:45.240]with a minimum of three prevacs
[00:01:47.680]for wrapped or difficult to penetrate items.
[00:01:51.740]Liquids cycle: this mode is used when autoclaving liquids
[00:01:56.690]to prevent boil over at the end of the cycle.
[00:01:59.540]Boil over is prevented by slowly releasing the pressure
[00:02:02.630]in the autoclave at the end of the run.
[00:02:05.260]Autoclave settings for solid materials
[00:02:07.330]release the pressure much quicker.
[00:02:10.450]Preparation of load: use only those types
[00:02:14.690]of containers, bags and lids
[00:02:17.080]that are designed for autoclaving.
[00:02:19.120]Inspect vessels for cracks or chips prior to autoclaving.
[00:02:23.040]Autoclave all contaminated wastes,
[00:02:25.150]including disposable plastic lab wear,
[00:02:27.470]for example culture plates, Petri dishes and pipette tips.
[00:02:31.690]Paper and gloves can also be autoclaved.
[00:02:34.710]Loosen lids on containers of liquids
[00:02:36.760]and closures on autoclave bags
[00:02:38.730]to allow for steam penetration.
[00:02:41.440]Always use secondary containment pans
[00:02:43.720]when sterilizing or decontaminating.
[00:02:46.320]Steel autoclave pans achieve appropriate temperatures
[00:02:49.460]quicker than polypropylene containers
[00:02:51.540]and thus are recommended.
[00:02:53.930]Use chemical indicator tape.
[00:02:55.870]It is very important to use a lead-free tape
[00:02:58.740]to avoid potential implications
[00:03:00.850]under the hazardous waste regulations.
[00:03:03.100]Consult EHS if you use an autoclave tape containing lead
[00:03:06.760]or if you are unsure if your autoclave tape contains lead.
[00:03:11.470]Preparation of load, sharps: autoclave sharps,
[00:03:16.410]for example razors, slides or scalpels
[00:03:19.120]only when contained within a rigid, leak-proof,
[00:03:22.000]puncture-resistant container.
[00:03:23.770]Make sure the container opening is set to the open position
[00:03:26.660]during the autoclaving process.
[00:03:28.750]Close the container opening after autoclaving.
[00:03:31.380]Be sure to deface any biohazard symbols
[00:03:34.070]with autoclave tape before autoclaving.
[00:03:36.950]This will show that the material is no longer a biohazard
[00:03:40.070]once the proper autoclave cycle is complete.
[00:03:43.840]Loading biohazardous waste into the autoclave:
[00:03:48.040]wear PPE such as lab coat, gloves and safety glasses.
[00:03:52.930]Ensure the biohazard waste bag
[00:03:55.010]is no more than three quarters full.
[00:03:57.750]Move the bag to a secondary container
[00:04:00.000]or autoclave pan positioned on a cart.
[00:04:02.770]Place an X with autoclave tape over the biohazard sign
[00:04:06.210]and loosely close the bag opening with autoclave tape.
[00:04:09.830]Move the load to the autoclave room.
[00:04:12.240]Slide the secondary container into the autoclave.
[00:04:14.760]Do not overload the autoclave.
[00:04:17.110]Seal the door and select the proper cycle for the run.
[00:04:20.810]Recommended settings for waste
[00:04:22.300]are at least 121 degrees centigrade
[00:04:25.840]at 15 to 17 psi for at least 30 minutes.
[00:04:29.610]Start the run,
[00:04:30.900]waiting to make sure the run starts correctly.
[00:04:34.460]Removing biohazard waste from the autoclave:
[00:04:38.810]ensure that the run is complete.
[00:04:40.630]Check the pressure gauge.
[00:04:42.000]If pressure is not released, do not open the door.
[00:04:45.320]Contact the Building Maintenance Reporter for malfunctions.
[00:04:48.590]Do not use the pressure relief override valve.
[00:04:51.840]Wear PPE, heat-resistant gloves,
[00:04:54.280]eye protection, lab coat and close-toed shoes.
[00:04:57.640]Have a cart ready.
[00:04:59.730]While wearing eye protection and insulated gloves or mitts,
[00:05:02.980]carefully open the autoclave,
[00:05:04.540]using the door to shield your body
[00:05:06.220]from the contents of the autoclave.
[00:05:08.480]Hot condensate may drip from the door,
[00:05:10.580]so ensure your feet are protected.
[00:05:13.410]Use caution when removing liquids.
[00:05:15.860]Liquids, especially large volumes,
[00:05:17.880]may continue boiling for a time after autoclaving.
[00:05:21.390]Slide a cart to the opening of the autoclave
[00:05:24.190]and pull the autoclave's secondary container onto the cart.
[00:05:27.680]Place the cart in a low-traffic area
[00:05:29.960]while additional cooling occurs.
[00:05:33.000]Disposal of autoclaved waste:
[00:05:36.370]liquids that have been autoclaved
[00:05:38.030]may be poured down the sink if all chemical components
[00:05:40.730]are listed on the EHS sewer disposal list.
[00:05:44.010]If the liquid contains chemicals
[00:05:45.540]that are not approved for sewer disposal,
[00:05:47.490]the vessel must be tagged for pickup by EHS.
[00:05:50.750]Non-toxic solids, including lead-free autoclave tape
[00:05:54.210]that do not contain any chemical constituents
[00:05:56.620]regulated under the hazardous waste laws
[00:05:58.810]or radioactive material may be disposed
[00:06:01.220]in the regular trash following autoclaving and demarcation.
[00:06:04.930]Seal the bag by taping or tying the bag shut.
[00:06:08.520]Place the autoclaved waste bag in a black, opaque trash bag.
[00:06:13.070]If any free liquids, such as condensate are present,
[00:06:16.440]add sufficient absorbent to the bag.
[00:06:19.114]Autoclaved bags will not be taken to the dumpster
[00:06:22.000]or disposed of by custodial staff
[00:06:24.150]unless in a dark trash bag.
[00:06:26.620]Users are responsible to purchase those bags.
[00:06:30.820]Autoclave performance checks:
[00:06:34.100]as part of a campus-wide program,
[00:06:35.940]autoclaves that are used to sterilize biohazardous waste
[00:06:39.660]are enrolled in the EHS Autoclave Testing Program
[00:06:43.110]to ensure that autoclaves are reaching proper temperature
[00:06:46.020]and pressure to sterilize biohazardous waste.
[00:06:48.950]EHS created this program to assist researchers
[00:06:51.880]in confirming proper autoclave function
[00:06:55.010]to avoid unintentional release
[00:06:56.690]of biohazardous waste materials
[00:06:58.470]due to a malfunctioning autoclave.
[00:07:01.110]This testing is needed because autoclave indicator tape
[00:07:04.020]does not provide decontamination effectiveness.
[00:07:07.240]Tape only indicates that the outside of the container
[00:07:10.030]came to temperature.
[00:07:11.250]It does not reflect time or conditions inside the load.
[00:07:15.286]Autoclaves enrolled in this program
[00:07:17.080]are tagged with a sticker.
[00:07:18.710]Orange labels are placed on autoclaves
[00:07:20.780]used to sterilize waste from BSL-2 labs,
[00:07:23.810]and white labels are placed on autoclaves
[00:07:25.810]used by BSL-1 labs.
[00:07:27.840]Each autoclave is assigned an identification number
[00:07:30.660]based on the room where it is located
[00:07:32.540]and the number of autoclaves in the room.
[00:07:35.210]EHS distributes test kits that contain the following:
[00:07:38.060]test organism ampule
[00:07:40.040]containing Geobacillus stearothermophilus,
[00:07:42.550]a shipping tube, documentation form and instructions.
[00:07:46.860]These kits are sent out at regular intervals
[00:07:48.960]depending on the containment level of the labs
[00:07:51.750]that utilize the autoclave
[00:07:53.040]for biohazardous waste decontamination.
[00:07:56.420]The testing procedure: open the envelope
[00:08:00.660]and remove the 50 mL conical Falcon tube
[00:08:03.330]and documentation form.
[00:08:05.160]If the packing material appears wet,
[00:08:06.990]contact EHS for a replacement ampule.
[00:08:10.320]Open the Falcon tube and remove the packing material.
[00:08:13.640]Remove the ampule and inspect for cracks.
[00:08:16.100]If you will be autoclaving the same day as receipt,
[00:08:18.940]hold the ampule at room temperature until use.
[00:08:21.670]If you'll be autoclaving a different day,
[00:08:23.700]store the ampule in a refrigerator.
[00:08:26.720]The testing procedure for dry loads:
[00:08:30.490]obtain a pipette tip storage box.
[00:08:33.250]Place the ampule on its side inside the box.
[00:08:36.440]Use autoclave tape to secure the ampule.
[00:08:39.030]Close the lid of the box and place the box under your load.
[00:08:43.180]For liquid loads, the testing procedure
[00:08:47.260]is to use a piece of string tied around the ampule,
[00:08:49.980]suspend it in liquid and secure it
[00:08:52.530]to the outside of the container with autoclave tape.
[00:08:55.960]The testing procedure for tabletop
[00:08:58.020]and top-loading portable autoclaves and sterilizers:
[00:09:02.660]obtain a 50 mL conical centrifuge tube.
[00:09:05.460]Place the ampule in the tube.
[00:09:07.260]Place one piece of autoclave tape over the mouth of the tube
[00:09:10.290]to prevent the ampule from falling out.
[00:09:12.500]It is important to allow for steam to enter the tube.
[00:09:15.490]Using another piece of autoclave tape,
[00:09:17.320]secure the tube to the bottom of the autoclave bag
[00:09:19.920]or simply place the tube under the biohazard bags.
[00:09:23.660]For plant and soil testing,
[00:09:26.830]either use a box to hold the ampule
[00:09:28.840]or use the Falcon tube as shown before.
[00:09:31.490]There are special considerations
[00:09:33.010]for waste soil sterilization.
[00:09:35.090]Refer to the EHS SOP, Autoclave Operation
[00:09:38.320]and Performance Testing.
[00:09:40.540]Testing of the autoclave: autoclave the materials
[00:09:44.830]using the appropriate cycle or settings.
[00:09:47.010]A typical waste cycle is 15 to 60 minutes
[00:09:49.510]at 121 degrees Celsius and 15 psi.
[00:09:53.780]Allow time for the autoclave to cool down
[00:09:55.830]and for pressure to return to atmosphere.
[00:09:58.230]Using insulated gloves or mitts,
[00:10:00.270]remove the load from the autoclave.
[00:10:02.540]Remove the pipette tip box from the bottom of the tray
[00:10:05.010]and open box to remove ampule,
[00:10:06.870]or remove cover from liquid container
[00:10:08.750]and retrieve ampule from the liquid.
[00:10:11.170]Allow the ampule to cool before placing ampule
[00:10:13.990]in a Falcon 50 mL conical tube provided by EHS
[00:10:18.440]and secure with packing material.
[00:10:20.430]The following items must be in the envelope returned to EHS:
[00:10:24.550]the completed documentation form,
[00:10:26.710]original or copy of the printout from the autoclave,
[00:10:30.510]the Falcon tube with test ampule and packing material.
[00:10:34.060]Place the return address label on the envelope,
[00:10:36.830]and place in the campus mail.
[00:10:38.526](energetic music)

Autoclave Operation & Performance Testing

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