NEW USERS INDUCTION PACKAGE

Bosch Mass Spectrometry Facility (BMSF)

Version 2016. 1.1Xiaosuo Wang

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

NEW BMSF USERS INDUCTION CHECKLIST

For a quick and easy induction to the Bosch Mass Spectrometry Facility (BMSF) ‐ please tick each of the following checklist boxes below as you complete each task. Please see the BMSF Officer for assistance should you require. Some of contents and forms can be found online. Please also visit: http://sydney.edu.au/medicine/bosch/facilities/mass‐spectrometry/index.php for more information Complete the BMSF registration form and agree on terms and conditions Obtain a New User Induction Package from BMSF Officer The package comprises 7 sections 1.PC2 training documents

o PC2 OGTR training – MFB o OGTR guidelines for certification of a PC2 laboratory Facility (version 3.2‐effective 1 March 2013) o OGTR guidelines for the transport, storage and disposal of GMOs (version 1.1‐ effective 1 September 2011) o OGTR guidelines for certification of an Animal Facility (version 3.2‐effective 1 March 2013) o PC2 OGTR Quiz

2.WHS related training documents specific to work performed at BMSF

o Health and Safety Risk Managemento Safe Work Procedure o Priority Work Health and Safety Risks at BMSFo Work Health and Safety (WHS) Action Plano The University WHS information and link

3.BMSF Workshop flyers for HPLC and / or MS 4.Policy and Agreement on the use of BMSF facility in addition to the BMSF registration form 5. Frequently asked questions 6. A starting protocol and analytical instruments induction checklist 7.Fee structure You are physically inducted by BMSF Officer in the lab and related area: locations of first aid box, fire extinguishers; knowing emergency contact, exit routes and assembly point, hot desks. You are also inducted to the analytical and general equipment needed for your project. You understand that you need to be trained prior to the use of any equipment for the first time. The training is arranged with BMSF Officer. Arrange the request ‘for a swipe card access’ to MFB with BMSF Officer if required

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

Equipment online booking password and the key pad code are given

BMSF email list subscription Signature page signed and returned to BMSF Officer DO NOT HESITATE TO COME AND TALK TO THE BMSF OFFICER IF YOU ARE UNCERTAIN ABOUT WHAT TO DO.

Section 2 Working with GMOs

Section 1 Guidelines For Safe Working Practices in MFB

PC2 Facilities

Sarah Walsh

Medical Foundation Building

University of Sydney

CONTENTS

Working with GMOs

- PC2 Practices for GMOs

- Your Responsibilities

- OGTR Requirements and Guidelines

- Institutional Biosafety Committee (IBC)

- Types of Dealings

- Compliance

- Transportation of GMOs

- Biohazardous Waste

- Spill Procedures

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Guidelines For Safe Working Practices in MFB PC2 Laboratories

- Your Responsibilities

- Lab Safety

- Training

- Biohazards

- Risk Groups

- Containment Levels

- PC2 Practices

- Containment

Section 1

Guidelines For Safe Working Practices in MFB PC2 Laboratories

Your responsibilities

It is your responsibility to seek information about health & safety risks, be involved in discussion about how these risk will be managed and take responsibility for your own actions.

Our Expectations

It is our expectation that you will:

› Follow established safe work practices

› Participate in relevant safety training

› Report hazards and incidents (including near misses)

› Cooperate with WHS inspections, audits and investigations

› Accept and respond to WHS advice and recommendations

› Not put yourself or others at risk of injury or illness.

Laboratory Safety – General Rules

› No eating, drinking or storing food inside laboratory areas.

› Minimum PPE for working in the laboratory (eg. closed non-absorbent shoes, laboratory gown or coat, safety glasses and disposable gloves.

- PPE should stay inside the laboratory and may need to be decontaminated (eg. Autoclaved) prior to cleaning.

- Gloves are not to be used to handle door knobs, phones or other shared equipment.

› Secondary containment must be used when chemicals, microorganisms or radioactive substances are moved outside of the laboratory.

› All laboratory waste must be disposed of in accordance with the University's procedures for the disposal of hazardous waste.

Working After-hours

› Afterhours work is restricted to low or medium risk work and supervisor approval must be attained.

› The completion of high risk work outside of core hours requires written supervisor approval, following review of a risk assessment . Specific risk controls and conditions may be applied to this work.

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Restricted Areas

Some areas of the University pose specific risks to health & safety (eg. laboratories and workshops). Access to these areas is restricted. The entrances to these areas will display Authorised Entry Only signage.

Only staff, students and contractors who have been inducted to the area are permitted to enter unsupervised. Anyone else entering the area must be under the direct supervision of the area supervisor.

Local inductions must include information about :

› The specific hazards in the laboratory and risk of exposure/incident

› Safety measures taken in the area (eg. procedures, safety equipment and PPE).

The standard Authorised Entry Only signage provides basic information about: •The hazards in the area •The safety precautions required when entering or working in the area •Contact details for the Area Supervisor •Emergency Contacts for the Building.

Laboratory Safety Training

› Working with Chemicals

› Bio-safety Training

› Radiation Safety for Laboratory Workers

› Local training: PC2/Working with GMO’s

› Training is open to both staff and research students. Enrol via learning solution

› http://sydney.edu.au/whs/activities/training.shtml

A variety of other information is available on the WHS Website

› http://sydney.edu.au/whs/

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What are Biohazards?

› Any organism or organism-derived material that poses a

Potential a risk to an individual, the community or the environment.

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Genetically Modified Organisms (GMOs)

Non-GMOs (microorganisms, human &

animal tissues, blood & body fluids, etc)

Risk Groups

› All organisms are organised by risk group according to the degree of risk they pose to an individual, the community or the environment.

› Risk Group 1(low individual and community risk) – a microorganism that is unlikely to cause human plant or animal disease.

› Risk Group 2(moderate individual risk and limited community risk) – a pathogen that can cause human, plant or animal disease but is unlikely to be a serious hazard to lab workers, the community, livestock or the environment. Lab exposure may cause infection but effective treatment and preventative measures are available.

› Risk Group 3(high individual risk and limited community risk) – a pathogen that usually causes serious human or animal disease and presents a serious hazard to lab workers. Could present a risk if spread in the community, effective treatment and preventative measures are usually available.

› Risk Group 4(high individual and community risk)- a pathogen that usually cause life-threatening human or animal disease. Presents a serious hazard to lab workers, readily transmissible from one individual to another. Treatment and preventative measures are usually not available.

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Physical Containment

› The level of facility with its practices and equipment must be suitable to work with the organism in question.

› Physical Containment Level 1(PC1)

Organisms from Risk Group 1 are used in this facility level.

These organisms are not know to cause disease in healthy adults. This could include organisms that have been inactivated or fixed.

› Physical Containment Level 2(PC2)

Organisms from Risk Group 1 & 2 are used in this facility level.

This is applicable to human and animal, clinical and diagnostic specimens.

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Signage

- OGTR PC2 sign and Biohazard Sticker (blue) at the entrance to all facilities.

- ‘No Food or Drink’ & ‘No flammable materials’ sign posted on all laboratory fridges and freezers.

- Outside the certified facility:

1. Biohazard sticker (black & yellow) on outside of all storage areas or devices that house GMOs - Only if storing Risk Group 2 GMOs.

2. Label area/device to indicate that GMOs are being stored.

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OGTR certified facilities require:

PC2 Practices

› Keep doors and windows closed at all times

› Do not bring personal items such as mobile phones or backpacks into the PC2 lab

› Take care that reading and writing materials do not become contaminated.

› Decontaminate work benches, bio-safety cabinets etc, after spills and when work is completed

› No eating, drinking, smoking or application of cosmetics

› No storage of food or drink

› Wear lab gowns and gloves and remove before leaving

› Wash hands with dedicated soap before leaving the lab

› You must wear closed-in footwear (foot enclosed).

› No mouth pipetting

› Take care when using sharp implements, never re-sheath needles – dispose all sharps directly into approved sharps containers

› Protect skin lesions and existing wounds prior to commencing work

› Don’t moisten labels with the tongue or chew pens

› Keep hands and pens away from your face. They may have been in contact with contaminated surfaces or aerosols

› Clean up all spills immediately

Personal Protective Equipment

› Lab coat or lab gown

› Gloves if working with Risk Group 2 micro-organisms

› Safety Glasses or › Safety Goggles (if there is risk of hazardous material splash to the eyes)

› Closed-in shoes

› Long hair tied back

› Cover all cuts and abrasions

› Face shield and cryo-resistant gloves if working with liquid nitrogen

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Footwear

› Footwear enclosing the entire foot must be worn when handling GMOs and when working in PC2 laboratory.

› Porous footwear such as sneakers should be avoided.

› Sandals, flip-flops or thongs are not permitted in laboratories.

› Socks with sandals is just plain wrong and should not be worn by anyone EVER!!

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Footwear worn within PC2 facility

Good Not Ideal

Not permitted - Fail Not permitted –Fail!

Containment Equipment

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ALL work that produces aerosols which may cause a significant risk to humans or the environment from the production of infectious aerosols must be performed in the biological safety cabinet.

Wipe down the internal surfaces of the BSC with (80% (v/v) ethanol) before and after use. Decontamination with UV light for 20 minutes is also an option. The biological safety cabinets are NOT fume hoods - They will not protect you from harmful chemical fumes.

Aerosols

› Aerosol hazard - breaking the surface of any liquid creates a cloud of aerosol droplets that can spread GMOs or microorganisms

› If your samples are biologically hazardous or infectious a Biosafety cabinet must be used while processing them.

› This applies to syringing, pipetting or dispensing samples, sonication, homogenizing, vortexing, centrifuging, shaking etc.

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Class II Biological Safety Cabinets (BSC)

› Training video: ‘Working safely in a Class II Biological Safety Cabinet’- http://www.vidrl.org.au/publications/biosafetycabinets/biosafetycabinets.htm

› Keep apparatus/material inside BSC to a minimum

› Check air pressure gauges for operational pressures before use,

› Don’t obstruct the air flow vents around work zone.

› After use: decontaminate work zone with 80% (v/v) ethanol and run cabinet for at least 5 mins.

› Changes in air pressure in the room will affect BSC operation (ie operator protection & biocontainment)

› BSC must be certified annually – before use check that certification sticker is in date.

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To operate a Biosafety Cabinet correctly:

Section 2 Working with GMO’s

› A GMO is defined as - An organism that has been modified by gene technology.

- An organism that has inherited traits from an organism that occurred because of gene technology.

- Anything declared by the Regulations to be a genetically modified organism, or that belongs to a class of things declared by the Regulations to be genetically modified organisms

- but does not include: • a human being, if the human being is covered by paragraph (a) only because

the human being has undergone somatic cell gene therapy.

• an organism declared by the regulations not to be a genetically modified organism.

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PC2 Work Practices for GMOs

– Store all GMOs in labelled sealed containers and ensure they are easily identifiable as a GMO.

– Access to storage (external to the certified lab) must be restricted to authorised personnel only, to prevent unintentional release of GMOs

– Dispose of all contaminated waste safely and in accordance with relevant SOPs refer to OGTR Transport, Storage & Disposal Guidelines.

– Wipe down ALL work surfaces before and after use with 80% (v/v) Ethanol

– Adhere to any specified license conditions as required by the IBC

eg. Volume limits for cultured GMOs, handling limitations for GMO animals (ie only in specified locations), specific researcher list for each GMO dealing etc.

– Remove both gown and gloves just before washing hands and leaving the facility.

– Always wash your hands with the antiseptic hand-wash provided before you exit the PC2 facility.

– Report all GMO spills to the Lab Manager and your Supervisor

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Personal Responsibilities

› Be familiar with the hazards and risks associated with the GMOs used in your lab.

› All lab work is to be carried out with regard to your personal safety and the safety of others in the lab.

› Be familiar with all relevant OGTR guidelines and training requirements:

› Use appropriate personal protective equipment.

› Follow safe work practices, standard operating procedures and policies.

› Participate in roster systems to maintain facilities such as tissue culture rooms. Keep all areas, including communal areas, clean at all times. Refer to specific procedures e.g. tissue culture.

› Report all incidents to the Laboratory Manager and your Supervisor.

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Office of Gene Technology Regulator (OGTR)

OGTR (mandatory compliance)

Gene Technology Act 2000: nationally consistent regulatory system for gene

technology

Gene Technology Regulations 2001: inter-governmental agreement

“for the regulation of genetically modified organisms in Australia, in order to protect

the health and safety of Australians and the Australian environment by identifying

risks posed by or as a result of gene technology, and by managing those risks

through regulating certain dealings with genetically modified organisms.”

Amendments to the Act and Regulations took effect on 1st July, 2007

New guidelines for certification took effect on 1st March 2013

www.ogtr.gov.au

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Office of Gene Technology Regulator (OGTR)

What does the OGTR do? › Administers the Gene Technology Act 2001 and Regulations

- monitors compliance with the Act and Regulations

- certification of facilities (PC1 to PC4)

- approves GMO Licenced Dealings

- DNIR (PC2 to 4)

- DIR (field release)

- regulates import and transport of GMOs

- investigates breaches in compliance

- can inspect a facility and associated GMO dealing at any time

- informs and advises other regulatory agencies and the public about GMOs and GM products

- publishes quarterly reports (including breaches)

- researchers deal with the OGTR through their IBC (Institutional Biosafety Committee)

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Institutional Biosafety Committee

› Any University staff or student wishing to undertake a project/dealing involving GMOs must apply to the University of Sydney Institutional Biosafety Committee (IBC) for approval

› IBC consists of expert staff members as well as a lay person.

› The IBC meets quarterly (meeting dates available on WHS website). Applications must be submitted at least 3 weeks prior to the meeting date to allow for applications to be photocopied and distributed to committee members for their consideration. All applications forms should be submitted to the University's Biosafety Advisor (Jenny Thomson).

› Further information regarding USYD IBC committee can be found at:

http://sydney.edu.au/whs/guidelines/biosafety/gene_technology.shtml

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How does the IBC help you work with GMOs?

› Report activities to the OGTR (annual)

› Annual inspections of certified PC2 facilities to monitor compliance

› Advice regarding the identification and management of risks associated with GMO dealings, including GMO containment

› Assessment & approval of GMO “dealings” applications (eg Exempt Dealings, NLRDs, DNIRs etc)

› Application Forms available on website (http://sydney.edu.au/whs/guidelines/biosafety/forms.shtml)

› Assessing research proposals and determining classification

› IBC meets quarterly to approve projects, work can not begin until your application has been approved by the IBC

› Report annually to the OGTR on research being conducted at the University

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What else does the IBC do?

• Dealings involving some high risk microorganisms are now regulated by the Security Sensitive Biological Agents (SSBA) Regulatory Scheme, as of January 2009

– National Health Security Act 2007

– http://www.health.gov.au/SSBA

• Please contact the IBC if you intend to start new experiments involving pathogenic organisms that are classified as SSBAs

– refer to list of SSBAs: http://www.health.gov.au/SSBA#list

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Types of Dealings All dealings must be approved by the IBC

› Exempt Dealings (exempt from major restrictions, not exempt from notifying OGTR or certification)

› Notifiable Low Risk Dealings (NLRD)

Licensed dealings:

› Dealings Not Involving an Intentional Release into the environment (DNIR)

› Dealings Involving an Intentional Release into the environment (DIR)

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Exempt Dealings

› Lowest perceived risk; research involving very well understood organisms and processes for creating and studying GMOs

› Requires PC1 containment

› Work must not result in the release of the GMOs into the environment

› No requirements to report to the OGTR

› Requires approval from the IBC › Please see 'Classes of Dealings' at :

http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/exemptdealclass-2 for more information and examples.

Notifiable Low Risk Dealings

› NLRDs are described in the Gene Technology Regulations 2001 and are dealings with GMOs that have been assessed as posing low risk to the health and safety of people and the environment provided certain risk management conditions are met. NLRDs must be:

› conducted by people with appropriate training and experience within a facility certified to either Physical Containment level 1, 2 or 3.

› assessed by the IBC and,

› transported, stored and disposed of in accordance with the Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs

› IBCs must prepare a record of assessment of each proposed NLRD. Copies of the IBC record of assessment must be provided to the person/organisation which requested the assessment for the NLRD. The person/organisation must be able to make a copy of the record of assessment available to the OGTR upon request, and must provide information to the OGTR as a part of each organisation's annual report. Please see 'Classes of Dealings' at : http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/exemptdealclass-2 for more information and examples.

Dealings Not involving Intentional Release

› Dealings NOT involving an intentional release of GMOs into the environment are dealings with GMOs in contained facilities that do not meet the criteria for classification as Exempt Dealings or Notifiable Low Risk Dealings. These dealings must be licensed by the Gene Technology Regulator (the Regulator). The contained facilities used for conducting DNIRs must be certified and typically range from PC2 to PC4.

› The appropriate level of containment is determined by the risk group classification of the wild type (non-genetically modified parent) organism as outlined in the Australian/New Zealand Standard (AS/NZS 2243.3:2010) AND the risk(s) identified for dealings with the specific GMO.

› In general, DNIRs consist of dealings with GM pathogenic organisms, or GM organisms containing higher risk genes from pathogens or genes that encode toxins or confer an oncogenic modification or immuno-modulatory function.

› Dealings with viral vectors can be classified in the DNIR, NLRD, and exempt categories, therefore guidance on the correct classification of contained dealings with viral vectors has been developed

› Please see 'Classes of Dealings' at : http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/exemptdealclass-2 for more information and examples.

Dealings involving Intentional Release

› Dealings involving an Intentional Release (DIR) of GMOs into the Australian environment are dealings with GMOs outside contained facilities.

› These can range from small scale field trials (limited and controlled releases) of GMOs to general/commercial release of GMOs

› Examples include:

• grow a GM crop as a field trial

• grow a GM crop commercially

• release a GM fish into a waterway

What do I need to do to comply with GMO legislation?

› Ensure the work you undertake involving GMOs or SSBAs has appropriate approvals from the OGTR or IBC.

- Speak to your laboratory supervisor/head about the scope and approval status of the experiments you are undertaking.

- Laboratory supervisors/heads carry ultimate responsibility for ensuring appropriate approvals are in place.

- Don’t hesitate to ask for advice.

› When in certified PC2 areas work in accordance with the OGTR’s PC2 Behavioural Requirements and relevant SOPs

- Refer to:

• Guidelines for Certification of a PC2 Laboratory Facility, V3.2 2013 • Guidelines for Certification of a PC2 Animal Facility, V3.2 2013 - Found at:

http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/certifications-1#pc2

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Transport of GMOs (includes GM waste)

› Follow OGTR Guidelines for Transport, Storage and Disposal of GMOs – version 1.1 (effective from 1 Sept 2011).

› The guideline details GMO containment, security, decontamination & preventing release or loss of GMOs.

› Denatured or fixed samples, that render the GMO as non-viable, are exempt

› Within the PC2 ‘facility’ use primary container

› Inside the building (eg between floors or PC2 facilities in same building)

- use secondary sealed container (e.g. Tupperware container)

- GMO waste contained in 2 unbreakable containers, one of which must be sealed (eg sealed bag inside secured wheelie bin)

› Outside the building or to other institutions:

- A primary container must be packed inside a sealed, unbreakable secondary container.

- GM micro-organisms or animals - must be wholly contained inside a sealed, unbreakable primary container and placed inside a sealed, unbreakable secondary container

- Label secondary container to indicate it contains a GMO & with name, address & contact details of sender. Include Biohazard sticker if Risk Group 2 GMO.

› Decontaminate outside of containers before transport & decontaminate all containers after transport

› Notify Lab Manager and Lab Head of GMO spills or loss outside the facility – must be reported to the IBC and in turn the OGTR

› Overseas or interstate transport must follow IATA, OGTR & AQIS requirements & rules.

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BIOHAZARDOUS WASTE

› All wastes containing or potentially contaminated with Genetically Modified Organisms (GMOs) or by-products derived from GMOs must be decontaminated by pressure steam sterilisation (autoclaving) or chemical treatment before collection and disposal by the waste contractor

› To ensure that waste is thoroughly decontaminated via pressure steam sterilisation, monitoring of the autoclave sterilisation cycles must be carried out in accordance with the autoclave guidelines.

› Wastes contaminated or potentially contaminated with GMOs that have been chemically decontaminated (eg with bleach) must be disposed of as chemical waste.

› Animal carcases that are contaminated with GMOs or infectious microorganisms must be autoclaved before disposal as biological waste.

› Bedding from animals that are infected with GMOs or infectious microorganisms must be autoclaved before removal from the cage and before disposal as biological waste.

› Any transfected cells should be treated as biological waste irrespective of being a GMO or not. If the cell line will be producing virus particles you will need the same level of decontamination as the parent virus would need.

› Refer to Hazardous Waste Disposal Guidelines – Clinical and Biological Waste: (http://sydney.edu.au/whs/guidelines/hazardouswaste/index.shtml)

Spills inside Biological Safety Cabinet

› Ensure cabinet remains operational

› Place absorbent material wetted with appropriate disinfectant over the spill (Bleach 1%)

› Allow 10 minutes to effect disinfection

› Disinfect gloved hands and remove protective gloves in the cabinet

› Remove clothing for sterilisation

› Put on a clean set of gloves and protective clothing for carrying out the remainder of the clean up

› After initial disinfection of spill, remove sharp objects with forceps and discard as contaminated sharps

› then remove excess fluid with absorbent material and discard into a container for sterilisation

› Wipe down the floor, cabinet work zone and remaining items of equipment with fresh disinfectant solution

› Disinfect both sides of the front grille and work floor within the cabinet

› Consider whether the cabinet should be decontaminated using either formaldehyde or vaporised sodium hypochlorite.

› The safety officer(Lab Manager) should be consulted for guidance

Spills outside a Biological Safety Cabinet

› In the event of a spill inside the laboratory, it should be assumed that an aerosol has been produced.

› Avoid breathing the aerosol.

› Warn others to leave the room immediately.

› Close the door and place a “DO NOT ENTER” sign on the door.

› Remove lab gown or other garment suspected of being contaminated, place in a biohazard bag for subsequent decontamination.

› If spill has soaked through laboratory clothing take a complete body shower.

› Notify the area supervisor and lab manager.

› Stay out of spill area for 30 minutes.

› Assemble a cleanup team consisting of three people, one to observe and direct, the other two to carry out the procedure

› Wear appropriate PPE – dedicated for spill clean up (eg gloves, gowns, safety glasses/goggles, P2 masks etc)

› Place absorbent material, wetted with the disinfectant (bleach 1%) over the spill, allow at least 10 minutes for disinfection

› Carefully remove any sharp objects with forceps and dispose of as contaminated sharps

› Clean up the spill and disinfectant solution

Spills OUTSIDE a Certified Facility

› Action exactly as you would a spill outside of a Biological Safety Cabinet (see previous slide).

› VERY IMPORTANT:

• Inform your supervisor, safety officer(lab manager) and IBC IMMEDIATELY

SUMMARY

› Understanding and complying with bio containment and biosafety requirements is everyone’s responsibility.

› If not sure – ask

- Supervisor/Lab Head

- Laboratory Manager (Sarah Walsh)

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Further Information

› Australian standards AS/NZS 2243.3:2002

› University of Sydney WH&S

- http://sydney.edu.au/whs/guidelines/biosafety/index.shtml

› Office of the Gene Technology Regulator

- http://www.ogtr.gov.au

› Laboratory Manager

- Sarah Walsh

- sarah.walsh@sydney.edu.au

- x63042

- 0423 295 788

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University of Sydney

Medical Foundation Building

PC2/OGTR Quiz

Training in compliance with:

- Australian Standards AS/NZ2243.3:2002

- OGTR Guidelines for Certification of a Physical Containment Level 2 Laboratory (V3.2 Effective 1 March 2013)

- OGTR Guidelines for the Transport, Storage and Disposal of GMOs (V1.1 Effective 1 September 2011)

- OGTR Guidelines for Certification of an Animal Facility(V 3.2 Effective March 2013)

PC2/OGTR Quiz Compulsory training for working in PC2/OGTR facilities at MFB

Questions to be answered from: PC2_OGTR Training SW 070415 V3 Section 1: Guidelines for Safe Working Practices in MFB PC2 Facilities Section 2: Working with GMOs Name: _____________________ Supervisor: _____________________ 1. What does GMO stand for?

_______________________________________________________________ _______________________________________________________________

2. What does OGTR stand for?

_______________________________________________________________ _______________________________________________________________

3. What does IBC stand for and what do they do?

_______________________________________________________________ _______________________________________________________________

4. List the four different types of dealings?

_______________________________________________________________ _______________________________________________________________

5. Do Exempt Dealings require approval from the IBC?

_______________________________________________________________ _______________________________________________________________

6. Briefly describe the decontamination procedure for a GMO.

_______________________________________________________________ _______________________________________________________________

7. Who must you notify if you have a spill of a GMO outside a certified facility?

_______________________________________________________________ _______________________________________________________________

University of Sydney

Medical Foundation Building

PC2/OGTR Quiz

Training in compliance with:

- Australian Standards AS/NZ2243.3:2002

- OGTR Guidelines for Certification of a Physical Containment Level 2 Laboratory (V3.2 Effective 1 March 2013)

- OGTR Guidelines for the Transport, Storage and Disposal of GMOs (V1.1 Effective 1 September 2011)

- OGTR Guidelines for Certification of an Animal Facility(V 3.2 Effective March 2013)

8. Describe the transport requirements when moving waste to the Animal Facility autoclave. _______________________________________________________________ _______________________________________________________________

9. Can you transport a GMO to a non-certified facility?

_______________________________________________________________ _______________________________________________________________

10. What is a biohazard?

_______________________________________________________________ _______________________________________________________________

11. What is a Risk Group 2 micro-organism?

_______________________________________________________________ _______________________________________________________________

12. List the risk groups of micro-organisms that can be used in the lab that you are

associated with. _______________________________________________________________ _______________________________________________________________

13. How often must eye wash and safety showers be checked?

_______________________________________________________________ _______________________________________________________________

14. Description of footwear permitted in the lab: _______________________________________________________________ _______________________________________________________________

15. Describe the PPE that must be worn when working with liquid nitrogen: _______________________________________________________________ _______________________________________________________________

Trainee signature: _________________________ Laboratory manager signature: _________________________ Date: _______________

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Faculty/Division: Sydney Medical School School/Unit: Medical Sciences /Bosch mass spectrometry Facility

Document number

Initial Issue date 05/2013

Current version V02

Current Version Issue date 02/01/2016

Next review date 02/02/2017

Risk Assessment name Handle flammable and Toxic solvents

Form completed by Xiaosuo Wang Signature

Date 18/02/16

Responsible supervisor/ authorising officer Paul Witting

Signature

Date 18/02/16

List legislation, standards, codes of practice, manufacturer’s guidance etc used to determine control measures necessary

Work Health and Safety Act 2011 Work Health and Safety Regulation 2011

AS 1940:2004 The Storage and Handling of Flammable and Combustible Liquids

Health and Safety Risk management form

Identify the activity and the location of the activity

Description of activity

Dealing with flammable solvents, such as ACN, MeOH, EtOH. Isopropanol

Description of location

G03A, medical Foundation Building, K25

Identify who may be at risk from the activity:

This may include fellow workers, visitors, contractors and the public. The types of people may affect the risk controls needed and the location may affect the number of people at risk

Persons at risk BMSF users, BMSF officer

How they were consulted on the risk

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Task or

Scenario

Hazard

Associated harm

Existing risk controls

Current risk Any

additional

controls

required?

Residual risk

Handling flammable and or /Toxic solvents ACN, EtOH, MeOH

Toxic/ Poison

Aerosols; Over exposure due to Inhalation may

result in irritation of the nose and throat, coughing, nausea and headache. Over exposure may result in dizziness, drowsiness, weakness, fatigue, breathing difficulties and unconsciousness. Ingestion may result in nausea, vomiting,

abdominal pain, diarrhoea, dizziness and drowsiness. Ingestion of large quantities may result in acidosis, visual effects, optic nerve damage, circulatory and respiratory collapse, coma and death.

Handle all solvents in the fume hood

Wear personal protective equipment: Wear splash-proof goggles, barrier or neoprene or PVC gloves and coveralls –lab coat. Where an inhalation risk exists, or respirator if necessary. Before use carefully read the product label. Use of safe work practices are recommended to avoid eye or skin contact and inhalation. Observe good personal hygiene, including washing hands before eating. Prohibit eating,drinking and smoking in contaminated areas.

Low No No

Handling flammable and or /Toxic solvents ACN, EtOH, MeOH

Flammable Fire risk; Solvents are highly flammable

Highly flammable. May evolve toxic gases

(carbon oxides, hydrocarbons) when heated to decomposition. Vapour may form explosive mixtures with air. Eliminate all ignition sources including cigarettes, open flames, spark producing switches/tools, heaters, naked lights, pilot lights, mobile phones etc. when handling. Earth containers when dispensing fluids.

Avoid heat, sparks, open flames and other ignition sources.

Low No No

Identify hazards and control the risks. 1. An activity may be divided into tasks. For each task identify the hazards and associated risks. Also list the possible scenarios which could sooner or later cause harm. 2. Determine controls necessary based on legislation, codes of practice, Australian standards, manufacturer’s instructions etc. 3. List existing risk controls and any additional controls that need to be implemented

3 | P a g e

Task or

Scenario

Hazard

Associated harm

Existing risk controls

Current risk Any

additional

controls

required?

Residual risk

Handling flammable and or /Toxic solvents ACN, EtOH,

MeOH

Irritant /inhalation

Contact may result in drying and defatting of the skin, rash and dermatitis. May be absorbed through skin with harmful effects. Over exposure may result in irritation of the nose and throat, coughing, nausea and headache. Over exposure may result in dizziness, drowsiness, weakness, fatigue, breathing difficulties and unconsciousness.

remove contaminated clothing and flush skin and hair with running water. Continue flushing with water until advised to stop by a Poisons Information Centre or a doctor If in eyes, hold eyelids apart and flush continuously with running water. Continue flushing until advised to stop by a Poisons Information Centre, a doctor, or for at least 15 minutes.

Low No No

Handling flammable and or /Toxic solvents ACN, EtOH, MeOH

Spill of solvents, Aerosols, as above Ingestion: as above. Highly flammable Contact with skin, eyes as above

Contact emergency services where appropriate. Use personal protective equipment. Clear area of all unprotected personnel. Ventilate area where possible. Contain spillage, then cover / absorb spill with non-combustible absorbent material (vermiculite, sand, or similar), collect and place in suitable containers for disposal

Low No No

Implementation Additional control measures needed: Resources required Responsible person Date of implementation

List emergency procedures and controls List emergency controls for how to deal with fires, spills or exposure to hazardous substances and/or emergency shutdown procedures Fire blanks, chemical spill kit.

4 | P a g e

REVIEW Scheduled review date: 12 months 2 years 3 years

Are all control measures in place?

Are controls eliminating or minimising the risk?

Are there any new problems with the risk?

Review by: (name)

Review date:

Risk management name and version number: I have read and understand this risk management form

Name Signature Date

Acknowledgement of Understanding

All persons performing these tasks must sign that they have read and understood the risk management.

Note: for activities which are low risk or include a large group of people (e.g. open days, BBQ’s, student classes etc), only the persons undertaking the key activities need to sign below. For all others involved in such activities, the information can be covered by other methods including for example a safety briefing, induction, and/or safety information sheet (ensure the method of communicating this information is specified here)

5 | P a g e

_______________________________________________________________________________________________________________ Page 1 of 2

Safe work procedure

Faculty/Division: Sydney Medical School

School/Unit: Medical Sciences /Bosch Mass Spectrometry Facility

Document number

Initial Issue date

Current version Current Version Issue date

Next review date

2.Safe work procedure title and basic description of activity

Title: Handling Flammable and Toxic Solvents Description of activity:- making up mobile phases for HPLC using flammable and toxic solvents

3.List Hazards and risk controls as per risk assessment

Associated risk assessment number and location: Risk assessment 001 BMSF (G03A), K25

Hazards As per risk assessment 001

Controls As per risk assessment 001

4.List resources required including personal protective clothing, chemicals and equipment needed Fume hood, Measuring cylinders,Glass pipette,Goggles, Nitrile gloves,Lab coat,Respirators 5.List step by step instructions or order for undertaking the task 1, Take the solvent bottle (flammable and / or toxic) from yellow solvents storage cabinet into fume hood 2, Open the lid of solvent bottle in the fume hood 3, Measure the correct amount of solvent in need using a correct size measuring cylinder i.e., 1L, 500 mL, 250 mL for various volumes of solvents 4, Transfer the correct amount of solvents into the screw cap HPLC solvent reservoirs /bottle (1L or 500 mL) 5, Put the solvent bottle back into yellow storage cabinet after ensuring the lid closed properly. 6, If the second solvent required, repeat the procedure from step 1. Ensure there is only one type of solvent present in the fume hood at a time 6, After completion of preparation of mobile phases /solvents, close the lid of HPLC reservoirs /bottles tightly before transferring them to HPLC solvents tray 6.List emergency shutdown procedures All solvents must be stored away from ignition source in case of emergency. All emergencies call 0-000; Display emergency procedures in area

7.List Emergency procedures for how to deal with fires, spills or exposure to hazardous substances Evacuate area and contact emergency services. Toxic gases may be evolved in a fire situation. Remain upwind and notify those downwind of hazard. Wear full protective equipment including Self Contained Breathing Apparatus (SCBA) when combating fire. Use waterfog to cool intact containers and nearby storage areas 8.List Clean up and waste disposal requirements Wearing the protective equipment outlined, ensure all ignition sources are extinguished. For small quantities, absorb on paper, sand or similar and evaporate under a fume cupboard or open area. For large volumes, atomise into incinerator (mixing with more flammable solvent if required) or recycle by gravimetric separation, distilling & reusing. Contact the manufacturer for additional information if required. 9.List legislation used in the development of this SWP AS 1940:2004 The Storage and Handling of Flammable and Combustible Liquids

1. Completed by: Xiaosuo Wang Staff/Student number: 1048792

_______________________________________________________________________________________________________________ Page 2 of 2

Occupational Health and Safety Act 2000 10a.List competency required – qualifications, certificates, licensing, training - e.g. course or instruction: All users using BMSF facility must have induction training (PC2 lab regulations) and instructions on safe use of the facility by BMSF officer 10b. List competency of Assessor 11.Supervisory approval, And review

Supervisor: Paul Witting Signature: Responsibility for SWP review: Sarah Walsh Date of review: 12.SWP Sign off sheet SWP name and version: Xiaosuo Wang Version 1.1 In signing this section the assessor/ authorisor agrees that the following persons are competent in following this SWP

Name Signature Date Competent

Name of Assessor/Authoriser

Assessor/Authoriser signature

The University of Sydney

Priority Work Health and Safety Risks

Faculty Sydney Medical School

School (if applicable) Medical Sciences

Discipline (if applicable)

Description of process or activity 1 Handle flammable and Toxic solvents

Location (campus, building, room other) Camperdown, K25, G03A

Name of manager in charge Dr Xiaosuo Wang

Phone Number 9036 3460

Email address Xiaosuo.wang@sydney.edu.au

Description of process or activity 2 Handle Gas Cylinders

Location (campus, building, room other) Camperdown, K25, G03A

Name of manager in charge Ms Sarah Walsh

Phone Number 9036 3042

Email address Sarah.walsh@sydney.edu.au

Description of process or activity 3 Chemicals brought in by users

Location (campus, building, room other) Camperdown, K25, G03A

Name of manager in charge Dr Xiaosuo Wang

Phone Number 9036 3460

Email address Xiaosuo.wang@sydney.edu.au

Description of process or activity 4

Location (campus, building, room other)

Name of manager in charge

Phone Number

Email address

Description of process or activity 5

Location (campus, building, room other)

Name of manager in charge

Phone Number

Email address

If you have many high risk WHS activities, select the “TOP FIVE”. Prioritise your “TOP FIVE” WHS risks by considering the likelihood and potential consequence of exposure and injury. For further guidance visit: http://sydney.edu.au/whs/activities/prioritise.shtml

WORK HEALTH AND SAFETY ACTION PLAN

Name of organisational unit and date of action plan: Bosch Mass Spectrometry Facility, 02/04/2013

Organisational units are required to set goals for achieving and improving performance in work health and safety (WHS). These WHS Action Plans should be established with reference to:

Local WHS risk profile and priorities

WHS reports and recommendations

The University’s WHS Action Plan.

How to use the WHS Action Plan template:

1. Tick the box for each action item that has an assigned action.

2. Note related details in the space below each goal.

3. Record “WHS Action Plan Summary” on Page 2.

WHS Goals 1. Risk management steps

X Identify the top five priority hazards or hazardous jobs and agree on a schedule for addressing these. X In priority order, assess each hazard and decide on suitable risk controls. X Implement agreed risk controls within designated time frame. X Schedule checking to ensure risk controls are working effectively.

Hazard Action By Whom By When Comments

Flammable and

Toxic Solvents

Training users to

ensure solvents are

handled safely

Users to follow

existing SOP

Write risk assessment

BMSF officer

Users

BMSF officer

As required

As required

April 2013

Gas cylinder use

and handling

Gas cylinder handling

training

Use of correct gas

Cylinder Trolley

Write risk assessment

Lab Manager

Users

Lab manager

As required

As required

May 2013

Chemicals brought

in to BMSF by users

BMSF officer should

always be notified of

any new chemicals

brought in to BMSF

Ensure all chemicals

are accompanied by

MSDS

Users

users

As required

As required

during induction to BMSF,

new users are notified of

this information

WHS Action Plan

WHS Services – 9351 4335 18 February 2016 Page 2

2. Hazard, incident and injury reports X Check reporting within 24 hours of occurrence or observation.

Check corrective actions are submitted (where relevant) within one week of notification.

Check corrective actions are completed on time and closed in RiskWare.

As yet there have been known incidence reported in BMSF.

3. Emergency preparedness X Allocate responsibility for coordinating emergency response. X Provide workers with training and practice in emergency procedures. X Provide after-hours emergency contact details to Security Services. X Assign responsibility for providing first aid service, or identify accessible first aid service nearby. X Communicate details of nearest first aid officer eg. persons name, phone, location. X Ensure emergency contact numbers are known ie., 9351 3333 for Security Service, 000 for fire, police or ambulance service.

BMSF officer provides training to users with proper procedures of handling flammable solvents.

Emergency contact details are placed on the door.

4. WHS training and promotion X Assign relevant WHS training for workers. X Ensure WHS induction is completed for new workers within their first week at work.

5. Monitor work practices and environment

X Conduct regular ie. quarterly walk-through WHS inspections of each work area.

Lab manager does walk-through WHS inspections of each work area of BMSF.

6. Contracts and services

Include relevant WHS issues in contract specifications.

Ensure contractors are qualified and informed of local hazards.

Provide induction and supervision.

Contractors are inducted by Brookfileds Multiplex Services.

7. Purchasing

X Assess hazards relating to equipment and materials being purchased eg. ergonomics, weight, noise, dust, heat, radiation, electromagnetic fields etc.

WHS Action Plan

WHS Services – 9351 4335 18 February 2016 Page 3

WHS Action Plan summary (As above ) Name Actions Due by Date

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

WHS AT THE UNIVERISTY OF SYDNEY

For more WHS related information, please go to the link below:

http://sydney.edu.au/whs/guidelines/index.shtml

The course utilizes a lecture and lab format at the Bosch Mass Spectrometry Facility (BMSF). Topics include general introduction for mass spectrometry, in focus of Triple quadrupole MS and its ionisation.It also covers HPLC columns and maintenance, data acquistion, worklists, MassHunter Qualitative software and hardware maintenance.

A course designed for beginners

or intermediates to enhance skills

in Agilent 6460 series Triple

Quadrupole LC/MS use.

UPLC-QQQ-MS WORKSHOP

BOSCH MASS SPECTROMETRY FACILITY

21 & 22 March, 2016

Rm 143, Medical Foundation Building (K25)

92-94 Parramatta Road

ndst

Find out more about contenthttp:/sydney.edu.au/medicine/bosch/facilities/mass-spectrometry/training/index.php

UPLC 2016.ai 1 22/2/2559 23:09:49

Bosch Mass Spectrometry Facility Room G03A, Medical Foundation Building K25

POLICY AND AGREEMENT ON THE USE OF BMSF

1. Before registration, you must discuss your project with the BMSF Officer to ensure that

your project is achievable at BMSF and that estimated costs are understood.

2. Users to provide account details to the BMSF Officer to enable usage fee transfer of

incurred facility costs.

3. Users are required to provide the reference papers and related literature and protocols to

BMSF Officer before method development.

4. Once the method is set up, BMSF is not responsible for any unexpected results / data

produced in the sample.

5. You must advise the BMSF Officer if they intend to use any other solvents and buffers for

any purpose rather than those listed below:

Solvents Buffers

Acetonitrile Minimum HPLC grade Formic acid ≤ 0.1%

Methanol Minimum HPLC grade Ammonium acetate ≤ 10 mM

Isopropanol Minimum HPLC grade Ammonium formate ≤ 10 mM

Water Minimum HPLC grade Acetic acid ≤ 0.05%

6. When working with biological samples, including cells, users must discuss sample

preparation steps with the BMSF Officer.

7. The BMSF provides service for new application set up and method development on either

co-authorship collaboration or fee based.

8. For postgraduate students, when more support is required, in addition to method

development, co-supervision by BMSF Officer can be arranged.

9. If misuse/inappropriate use of equipment occurs and causes damage, users will be

responsible the repair cost incurred.

10. For online bookings, when using ESI ion source, users are allowed to book a maximum 2

consecutive days, if more booking days are required, the BMSF Officer must be contacted.

For APCI ion source, up to 5 consecutive days can be booked, but on a once per month

basis only.

11. If bookings are cancelled with less than 24 hours notice, $50 fine will be charged to

respective user’s account.

12. If users do not use the instrument within the first 2 hours of booking, the booking will be

cancelled with $50 fine.

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

FREQUENTLY ASKED QUESTIONS:

Do I need to purchase my own column?

Yes. We recommend that you purchase your own column if you have many samples (> 50

samples) to analyse, including matching guard columns.

The facility can also lend you a column, however there will be a one-off “loan” fee for this

service ($400).

Do I need to bring my own solvents and consumables?

No. We provide all necessary commonly used solvents and consumables, including general

consumables such as HPLC bottles, vials, caps, gloves, pipette tips, Eppendorf tubes, etc.

These consumables form part of the charge schedule per injection.

NB. We do not provide any special consumables such as filter vials, solid phase extraction

cartridges, etc.

Can I wash my column at the BMSF?

Yes, we can help you to set up a wash program on our instrument. This is a free service; a

wash step /method can be incorporated at the end of a run so that the column can be

stored safely.

Can I get help on method development and set up?

Yes. The method set up is either fee based or on the basis of a co-authorship agreement

between the researchers and the Facility Officer. The Academic in Charge (A/Prof Paul

Witting) is available for obligation free advice.

Can I prepare samples at the BMSF?

Yes. We have a range of general equipment available for sample extraction and sample

preparations including solid phase extraction and evaporation manifolds.

Are there any other costs involved in addition to the injection charge?

In addition to a small percentage of maintenance fee (usually 5% of total injection fee),

there are no other extra costs involved other than the injection charge when running

samples. There are however cancellation and damage exceptions as follows:

A cancellation penalty fee of $50.00 may be charged if a booking cancellation is given

less than 24 hours prior to the confirmed booking time.

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

The user will be responsible for any damage to instruments if deemed to be caused

by the misuse / inappropriate use. The repair cost will be charged to the user’s

account.

How to acknowledge the Facility in publications?

All research publications, either collaborative or non-collaborative, that contain data

obtained via resources at the BMSF are required to explicitly acknowledge such – example

as follows –

The authors acknowledge the support received from the Bosch Institute's Mass

Spectrometry Facility, and the expert help of Facility staff, insert name/s........

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

A STARTING PROTOCOL FOR LC-EC/UV/FL, LC-SQ, AND LC-QQQ-MS

Starting Checklist

The following items should be checked EVERYTIME BEFORE starting the system:

There is sufficient mobile phase for the expected analyses. Updating solvents (Line A, B, C,

and D) levels on the computer whenever new solvents are added. Always tick the boxes of

‘prevent analysis if solvent levels below 0.1 L’ and “turn pump off if running out solvents’.

o Do not refill any solvents while the instrument is running.

Turn pumps on (QQQ LC/MS system can be trigged on by clicking “purge the system”).

Purging the system:

LC-UV/EC/FL : 5 mL /min for 5 min

SQ LC/MS: 5 mL/min for 5 min

QQQ LC/MS: 10 mL/min for 5 min

o Remember to open purge valve for LC-UV/EC/FL and SQ LC/MS BEFORE purging. If

both separation (1260 pump) and make up (Bin pump) pumps used in SQ LC/MS,

they should be all purged.

o Do not over tighten purge valve when closing.

After purging, all lines of solvents should be bubble free, if any bubbles detected, re-purge

the line again till the bubble is removed.

The system should be running with last saved method after purging. Attaching one end of

your column first and watch the solvents flowing through and then connect the other end.

Slow down the flow rate if needed while connecting.

All connections / fittings are leak free. In most cases, connections should be finger tighten

only. Do not over tighten any connections.

For SQ LC/MS, trace the flow path to ensure the correct modules are connected when either

using MS only, or using active splitter for both fraction collector and MS analysis.

Turn other modules on, i.e., CCT, ALS, UV, MS, etc..

Open or edit your desired method and work through the entire method to ensure everything

is correct.

Column compartment and auto sampler temperatures are correct if applicable.

Monitor pumps pressure change. If it’s significantly different from your previous run, it’s

either blockage somewhere in the flow path (causing high pressure) or leakage in the

connections (causing pressure drop).

Start your run after carefully checking your method, workslist / sequence, sample

information, data storage path to ensure there is no mistake

o Start the run when all modules are green.

o Do not start the run when any of the modules is yellow or red.

Fill out the log book

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

BMSF PC2 LABORATORY INDUCTION CHECKLIST* ---- An addition to “A Starting Protocol”

Laboratory Access:

Registered users only;

PC2 lab inductions required;

Personal Protective Clothing and Equipment (PPCE) required.

Laboratory Practice

New applications need to be discussed with BMSF Officer to ensure the required analysis is

achievable within BMSF; the solvents used and sample preparation procedures are

compatible to normal performance of the instruments. It is imperative consult the BMSF

Officer when starting a new protocol.

Before you start, do a visual check on following items:

o The flow path connected to the desired detectors.

o If waste drum is full, please cap it and replace it with an empty drum located in the

second room G35; please ensure all waste lines go into the drum.

o If a solvent spill is seen on the ground, benches, and instruments please clean with

paper towel and note down in the log book; if a large spill is spotted, please notify

BMSF officer (Xiaosuo Wang) or Lab manager (Sarah Walsh +61 423 295 788)

If column needs to be changed, carefully remove the column and ensure the removed

column is CAPPED WELL on both ends. The previous user is responsible for storing the

column into an appropriate solvent.

Always use “HPLC grade solvents” that are supplied by the BMSF, measuring cylinders are

available in the fume hood to measure correct amount of solvents to prepare the mobile

phase. Always measure different solvents separately.

Cover the cylinder to protect from dust contamination with aluminium foil after use. The foil

is located on the shelf above the Eppendorf centrifuge.

Do not add any buffers or additives into the solvent measuring cylinder, add the buffers

/additives into the HPLC reservoir (bottle) directly instead.

Each instrument is allocated with a number of solvent bottles sitting in a plastic tray with

labels. Do not mix solvent bottles among those instruments.

If new clean bottles required, please ask BMSF Officer, if not available, please following the

below instructions to wash bottles:

o Discard any mobile phase from previous users into an appropriate waste bottle

/drum;

o Rinse the bottle with hot Millipore water (60 °C), repeat 3 x times, then rinsed with

HPLC grade Methanol once, Isopropanol once, and finally the running solvents

(mobile phase).

o In the case of hot water not available, use Millipore water 1x

o HPLC grade isopropanol 1x

o Millipore water 3 x

o Isopropanol 1x

o The solvents to be made up 1x

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

Note: Each time, vigorously shake /swirl to ensure all inside surfaces are in contact with

solvents.

Any new column or used column to be used at BMSF for the first time it needs to be washed

thoroughly prior to use with samples. Please following the below steps to wash your column

(assume they are all reversed phase):

o Fast wash: Create a two-hour gradient with optimal flow rate (based on column

dimensions. Please ask BMSF Officer if not sure) using solvent A: HPLC water;

solvent B: 90% Acetonitrile; the gradient starts from 0% B ramping up to 100% B

within 90 min and maintained at 100% for 20 min, and then decreased to 60%

within 2 min and maintained at 60% B till the end of the gradient.

o Alternatively, an overnight slow wash can be done (it’s also recommended when

time allows)

o Overnight slow wash: Using the same solvent, create a gradient with a very low flow

rate i.e., 0.1 mL/min for an analytical column. The gradient starts from 0% B ramping

to 100 % within a few hours and stayed at 100% for at least 2 hours before

decreasing to 60% B and maintained until the end of wash cycle.

If the column has been used in other instruments and intended to be used on QQQ, please

ask BMSF Officer before you start using it on QQQ

When using Single Q MS, before turning it on, the ion Source needs to be cleaned with 50%

isopropanol using Kimwipes. Be careful not to touch the nebulizer.

o Please do not clean QQQ’s ion source

Injection concentrations: for QQQ, it’s recommended to start with nM range and gradually

increase the concentration if the intensity of signal is not satisfied.

When running in a sequence or work list for multiple samples: blanks, system flush, column

storage methods need to be incorporated into the sequence / work list.

Ensure “Instrument Standby” script inserted at the end of run.

Always fill in the log book

Any enquiry, please contact Dr Xiaosuo Wang

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

FEE STRUCTURE AT BMSF The fee structure and conditions are listed below effective from 01-01-2016.

Descriptions:

Free access to all equipment housed at BMSF, provided that users are trained by BMSF

Officer;

Free training workshop and free on-site personal training;

Free consultation for column and SPE cartridge selections, including helping choose a

column and matching guard columns to ensure it’s appropriate for the application; obtain a trial

period for the column purchased;

When more support and guidance needed for postgraduate students, co-supervision shall be

arranged;

The fee is based on per injection charge regardless of number of analytes in the sample. In

the case of one sample needs to be analysed twice, this will be considered as 2 injections;

Users are responsible for special consumables such as cell culture medium, significant use of

N2 gas, SPE cartridges, filter vials.

The fee listed covers general consumables including solvents.

The fee is not based on the hourly usage.

Maximum $12,600 injection charge is applied through a calendar year. If more injections

occur after $12,600 value, there is no further charge.

Instrument/configuration/application/Items Fee ($ per injection)

Agilent HPLC 1200-UV

Agilent HPLC 1200-FL

Agilent HPLC 1200-EC

$6

$6

$8

Agilent HPLC 1260-UV/SQ MS $10

Agilent UPLC1290-UV/QQQ MS $12

Loan of column (without guard column) $400 (one off charge)

Service Maintenance Contribution 5 % of injection fee (i.e., 50c for a $10

injection )

Column wash Free

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

Column regeneration

Column maintenance

Free

Free

HPLC solvents, buffers, bottles, vials, inserts and

caps;

General consumables including tips and

Eppendorf tubes, etc.

Free

Method development On application or authorship agreement

Bosch Mass Spectrometry Facility Room G03A, Medical foundation Building K25

SIGNATURE PAGE

I have completed New Users Induction Checklist. I have read and agreed on the policies and

conditions listed.

Name:

Signature:

Date: