OHS026
Safe Work Procedure
Faculty/Division
Faculty of Medicine
Document number
School/ Divisional Unit
POWHCS
POWCS.RESP.SWP03
Initial Issue date
28/4/2010
Current version
1.2
Current Version
Issue date:04/04/2013
Next review date:
04/04/2014
The Writing Safe Work Procedures Guideline (OHS027) should be consulted to assist in the completion of this form.
Safe Work Procedure Title and basic description
Title: Exhaled breath condensate collection
Description: Collection of exhaled breath condensate of research participants
Associated risk assessment title and location:
Describe the activity or process
Preparation
1. Fill esky and thermos flask with wet ice.
2. Prepare mouth pieces by cutting two holes in one piece and one hole in another
3. Place curved glass condenser in test tube then place test tube in thermos flask
4. Insert mouth piece to end of curved condenser
5. Insert cloth/tissue barrier on top of thermos flask
Collection of sample
1. Obtain consent
2. Fill in subject data sheet
3. Ask patient to rinse mouth with water
4. As patient to conduct tidal breathing through the mouthpiece for 10-20 mins until ~1mL sample collected
5. Pause procedure if patient is dizzy, complains of dry mouth or excessive saliva
Processing of sample
1. Pipette 120uL into 6 or more eppendorf tubes
2. Place samples in esky with wet ice
3. Dearate condensate by introducing argon gas (0.4L/min) above sample inside eppendorf tube, maintaining for 20 seconds
4. Repeat step 3 for all eppendorf tubes, then place in esky
5. Place samples in freezer (-80C)
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Page 1 of 3
Safe Work Procedure
Uncontrolled document when printed
Date Effective: 01/01/2007
Current Version: 1.2,
04/04/2013

List all resources required including plant, chemicals, personal protective clothing and equipment, etc patient consent form
- subject data sheet
- cardboard mouthpiece with one-way valve
- non-siliconised glass condenser (at 4C)
- test tube
- wet ice
- cloth barrier
- thermos flask (cooling chamber)
- esky (-14C)
- pipette (120 uL)
- eppendorf tube x10
- pH meter
- argon gas
- milliQ water
- kidney dish
- miscellaneous (marker pens, scissors, paper tissues)
List potential hazards and risk controls including specific precautions required
1. Infection with diseases carried in patient saliva
Risk control: patient asked to disclose any recent illnesses in patient questionnaire; hand-washing before and after sample collection ; mouthpieces are disposed of after one use, glass components of collection device (condensing tube and test tube) washed after one use
2. Freezer burn
Risk control: wear cotton-padded gloves covering all of forearm when placing sample into freezer
List emergency shutdown instructions
Stop collection immediately
List clean up and waste disposal requirements
Waste disposal of mouthpieces – disposed into domestic waste bin (?)
Clean up procedure for glassware:
1. Depending on the amount of equipment, using a measuring cylinder, pour either i) 18ml or ii) 30 ml of Endozyme solution (formulation of protease and amylase, digesters and buffers) into the yellow tub.
2. Fill up the yellow tub with warm water to i) 3L or ii) 5 L mark. Obtain the warm water from the biohazard room. Soak the apparatus in the solution for 30 minutes.
3. Rinse the apparatus with Milli Q water from biohazard room.
4. Place test tubes in rack and fill with alcohol (70%). Place stoppers at one end of tubing and fill with alcohol (70%), placing it in standing positions (dependent on the long axis). Leave both test tubes and tubings for 30 minutes.
5. Drain the alcohol out and rinse it with Milli Q water 3 times.
6. Leave to dry on the bench top, covered by Readiwipe (cello paper/multipurpose cloth).
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Page 2 of 3
Safe Work Procedure
Uncontrolled document when printed
Date Effective: 01/01/2007
Current Version: 1.2,
04/04/2013

List legislation, standards and codes of practice used in the development of the SWP
N/A
Supervisory approval, training, and review
Supervisor: Paul Thomas Signature:
Plant custodian: Paul Thomas Signature
List competency required – qualifications, certificates, licencing, training - eg course or instruction: instruction
SWP review date: 23/4/2013 Responsibility for SWP review: Paul Thomas
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Page 3 of 3
Safe Work Procedure
Uncontrolled document when printed
Date Effective: 01/01/2007
Current Version: 1.2,
04/04/2013