safe working procedures HPLC 012


OHS026 Safe Work Procedure



School/ Divisional Unit

School of Medical Sciences

Document number SOMS GBD SWP 012 Initial Issue date


Current version


Current Version Issue date


Next review date


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


Use of HPLC

Description: Make peptides – radioligand compounds Associated risk assessment title and location: disposal of waste. RA Radiation spills 013, RA Waste management 015 located in Room 209B.

Describe the activity or process

HPLC Pumps  Fill the solvent reservoirs with sufficient solvent. If the HPLC system has not been used for long time, methanol should be used first to remove air bubbles from the solvent supply line and column.  Prime pump A and B from the draw – off valves with a syringe containing hypo needle   Turn the pump power switches to ON Flush the pumps for 15 min or longer at the flow rate of 1.0ml/min or longer for each pump depending on how long the system has not been used  Model 660 solvent programmer  Turn the thumbwheel switches to 0.0ml/min, if the pumps are driven by ‘MODEL 600 SOLVENT PROGRAMMER’. Set INITIAL CONDITIONS (pump B), FINAL CONDITIONS (pump B) HOURS: MINUTES and MILLELIERS/MINUTE (pump A&B) by turning the thumbwheel switches to desired values   Turn the power switch on by depress the power pushbutton Depress INTIAL CONDITIONS pushbutton on the left panel. Let the system run at the initial conditions for 15min, before switch on the ABSORBANCE DETECTOR (refer below). A further 30min at the initial conditions is required to warm up the detector.  Before injecting sample, make sure the HPLC injector LOAD/INJECT handle is at LOAD position. Fill a 1ml syringe with 0.5ml sample and inject the sample into the injector through an inject needle  Absorbance Detector (series 441)  Turn the LOAD/INJECT handle to INJECT position. At the same time depress RUN pushbutton on the left panel of Model 660. The program will start. When the program is finished, depress REVERSE PROGRAM button and the system will reverse from final conditions to initial conditions. Set the DIGITAL PANEL METER SELECTOR SWITCH (a knob on the top of right hand side) to ABS UPPER CH position   Set the record sensitivity to 0.5 by depressing the RANGER SELECTOR PUSHBUTTON Depress the +/- pushbutton and  1 pushbutton  Fraction collector (RETRIEVER 500)  Depressing POWER pushbutton to ON, warm up the UV light for 30 min before injection Set the MODE panel to desired value (min or sec) by pressing the BLUE button    set the DISPLAY panel to UNITS IN TUBE by pressing the BLUE button Put the racks onto the top of the collector and insert 5ml polypropylene tubes into the racks Insert HPLC outlet tubing into the HOLE of the BLACK SCREW STAND on the left side of the collector and adjust the stand to make sure the fluid dripping into the polypropylene tubes  Chart Recorder (RIKADENK1)  Press RUN/STOP (red button) to start or stop. Change tubes at anytime by pressing NEXT button Switch POWER button to ON   Set the CHART SPEED CONTROL to 12 cm/H and the AMPLIFIER to 2 Turn the CHART button to on ___________________________________________________________________________________________________________ ___________ Page 1 of 2 Safe Work Procedure Uncontrolled document when printed Date Effective: 01/01/2007 Current Version: 1.2, 15/08/2007

List all resources required including plant, chemicals, personal protective clothing and equipment, etc

  Suitable PPE Methanol, buffer, sample tubs, collection tubes, racks, radiation protect screen

List potential hazards and risk controls including specific precautions required

   Potential electrical hazard – annual inspection of electrical equipment and ensure inspection tag is current. Potential aerosol hazard – wear PPE, collect sample in fume cupboard. Spill of radioactive solution – Train all users to handle the radioactive solution in a right way, wipe the spill area with count off immediately and check working area using radiation monitor after finished work.

List emergency shutdown instructions

 HPLC has “OFF” switch that can be used in emergency.  Shut all electrical equipment off at power point in case of emergency.

List clean up and waste disposal requirements

   Dispose non radiation contaminated gloves and tubes in biological waste. Dispose radiation contaminated gloves and tubes in radiation waste and mark the level of radioactivity that have been disposed. Wipe all radiation contaminated area with count-off and monitor the working area every time when you have finished

List legislation, standards and codes of practice used in the development of the SWP

NSW OHS Act 2000 NSW OHS Regulation 2001 Australia Dangerous Goods Code Code of Practice for the Labeling of Workplace Substances AS/NZS 2243.2:2006. Safety in laboratories. Part 2: Chemical aspects Australian Standard AS2243.6-1990. Safety in laboratories. Part 6: Mechanical Aspects. Australian Standard AS2243.7-1991. Safety in laboratories. Part 7: Electrical Aspects. AS/NZS 2161.1:2000 Occupational Protective Gloves – Selection, Use and Maintenance Safe Work Procedure Form (OHS026) UNSW Hazardous Waste Disposal Procedure

Supervisory approval, training, and review

Supervisor: Prof E Burcher Signature: Plant custodian: Signature List competency required – qualifications, certificates, licensing, training - e.g. course or instruction: SWP review date:


Responsibility for SWP review: Fei Shang ___________________________________________________________________________________________________________ ___________ Page 2 of 2 Safe Work Procedure Uncontrolled document when printed Date Effective: 01/01/2007 Current Version: 1.2, 15/08/2007