OHS026 Safe Work Procedure - PDF

Document Sample
OHS026 Safe Work Procedure - PDF Powered By Docstoc
					OHS026

Safe Work Procedure

Faculty/Division Medicine                                            School/ Divisional Unit St George Clinical School

Document number            Initial Issue date          Current version             Current Version             Next review date
STGCL.SWP.4.1              15/09/09                    1.0                         Issue date 15/09/09         September 2010

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: Safe Operation of Hermle Benchtop Centrifuge Z 513 K
Description: This Safe Work Procedure describes the safe use & maintenance of Hermle Benchtop Centrifuge Z 513 K
Associated risk assessment title and location: STGCL.RA.4.1

Describe the activity or process
This procedure describes the preparation of samples for centrifugation, selection of rotor, selection of adaptors, how to position the
tubes in the adaptors to achieve balance prior to each spin, safest way to set up and operate the centrifuge, and appropriate
maintenance of this machine.

Preparation of samples for centrifugation
    1. Follow the relevant protocol for sample preparation to the stage where centrifugation is required.
    2. Select both the rotor and the tube type that you intend to use for the spin. The selection of rotor/tube will depend on the
         speed of the spin required, the volume of sample to be spun and the type of sample to be spun.

          Rotor Type Required
          There are two rotor options available with this centrifuge:
          Fixed angle rotor 220.70 V06:
          This rotor has a maximum speed of 21,000 x g (15,000 rpm) and has capacity for 8 x 50 ml round bottom tubes.

          The 50ml Blue Greiner centrifuge tube can also be spun in this rotor to a maximum speed of 9500 x g and no adaptor is
          required

          Swing Out Rotor 220.70 V06
          This rotor has a maximum speed of approx 7,000 x g (5,000 rpm) and has the following capacity:
          50 ml Blue Greiner centrifuge tubes in appropriate beige conical adaptor
          15 ml Blue Greiner centrifuge tubes in appropriate beige conical adaptor
          250 ml Nalgene Centrifuge bottle in beige flat bottomed adaptor

          Tube Type Required
          The tubes selected need to fit the rotor, be able to withstand the speed, be compatible with the chemicals being spun and
          be a suitable volume.

 Tube Type                   Minimum               Rotor           Adaptors Required /            Chemicals       Suitable Speed
                             Volume                                Type                           Resistant       (for this tube
                                                                                                                  type)*
 Nalgene Oakridge            40 ml (80% Full)      Fixed Angle     None required                  Very Good       Up to 50,000 x g*
 Round Bottom Tube                                 220.70 V06
 (Polypropylene
 Copolymer)
 Nalgene Centrifuge          200 ml (80% Full)     Swinging        Use Beige Adaptors             Very Good       Up to 13,200 x g*
 Bottles (Polypropylene                            Bucket Rotor
 Copolymer)                                        220.80 V02
 Greiner Centrifuge          50 ml Max Vol         Swinging        Use Beige Adaptors of          Very Good       Swing Out Rotor:
 Tubes (50 ml Blue Top       0.1ml Min Vol         Bucket Rotor    correct size                                   3200 x g
 Conical–Polypropylene                             220.80 V02
 Tubes with HDPE
 Caps)
 Greiner Centrifuge          15 ml Max Vol         Swinging        Use Beige Adaptors of          Very Good       Swing Out Rotor:
 Tubes (15 ml Blue Top       0.1ml Min Vol         Bucket Rotor    correct size                                   4000 x g
 Conical- Polypropylene                            220.80 V02
 Tubes with HDPE
 Caps)

          *Although the tubes are rated to spin to 50,0000 x g and 13,200 x g, these speeds cannot be reached with these centrifuge
          / rotor combinations because the maximum speed that the rotors can spin to are 21,000 x g for the Fixed Angle Rotor and
          7,000 x g for the Swinging Bucket Rotor.




___________________________________________________________________________________________________________
                                                          Page 1 of 5
Safe Work Procedure                                                                   Date Effective: 01/01/2007
Uncontrolled document when printed                                               Current Version: 1.2, 15/08/2007
    3.   Install the correct rotor as follows:
         For both rotors, use the Step available in the Lab to allow for easy access to the centrifuge. Also take care when lifting the
         rotors because these can be heavy.

         Installation of the Swing out Rotor: Check that the pin is located centrally and that the pin is straight prior to installing the
         rotor. If the pin is bent do not use the centrifuge and contact a member of Dept of Medicine Lab so that they can
         contact the service technician for replacement pin (contact details attached to the machine). Place the rotor on the
         motor shaft ensuring the pins align correctly with the rotor slots. Hold the rotor with one hand and secure the rotor to the
         shaft by turning the rotor nut counter clockwise (use the tool provided).

         Installation of the Fixed Angle Rotor:
         Check that the pin is located centrally and that the pin is straight prior to installing the rotor If the pin is bent do not use
         the centrifuge and contact a member of Dept of Medicine Lab so that they can contact the service technician for
         replacement pin (contact details attached to the machine). Place the rotor on the motor shaft ensuring the pins align
         correctly with the rotor slots. Hold the rotor with one hand and secure the rotor to the shaft by turning the rotor nut counter
         clockwise (use the tool provided).




         Fig 1: Balanced Swing Out Rotor                                                     Fixed Angle Rotor


    4.   Load Rotors with adaptors / tubes so that they are balanced
         See Figure 1 for examples of correct configurations of adaptors in the Swing Out Rotor.
         For Fixed Angle rotor, load tubes of equal weight opposite to each other – use a 1 decimal place balance to check
         that the weights of tubes placed opposite to each other are the same.

         For Swing Out Rotor, make sure to load the complete set of tube racks and buckets. If a combination of different tube
         racks & buckets is to be used:
              •    the same tube racks must be loaded opposite each other so that the load is balanced.
         Tubes placed OPPOSITE to each other MUST BE BALANCED ie EQUAL WEIGHT & IN RIGHT POSITION.
         Use a 1 decimal place balance to check that the weights of tubes placed opposite to each other are the same.
              •     –Some EXAMPLES of correct configuration are illustrated below.




              •     the tube racks must require the same radius correction, therefore tube racks for 50ml Blue Top Conical
                    Tubes and tube racks for 15ml Blue Top Conical Tubes CANNOT be spun at the same time. For further
                    information regarding the Radius Correction Factor, see section on Adjustment of the radius correction
                    factor.

         If spinning biohazardous material secure the hermatically sealed large transparent Lid over the Swing out rotor by pressing
          the snap connectors together on the rotor nut, pressing down the lid firmly down and then releasing the snap connectors.




___________________________________________________________________________________________________________
                                                          Page 2 of 5
Safe Work Procedure                                                                   Date Effective: 01/01/2007
Uncontrolled document when printed                                               Current Version: 1.2, 15/08/2007
     5.   Set Up and Operation of Centrifuge
                                                                          Knob 4 – Speed
          Power Switch                                                    adjustment
                                                                          increments of
          The main power switch is located on the left hand side of the centrifuge. When this switch is on, the digital display will be lit
          up.                                                             500

          Lid Release & Lid Lock
          The light on the lid button will appear green when the centrifuge can be opened safely. Press the Lid button and the
          centrifuge lid will open automatically.

          When the rotor has been inserted and secured correctly (see point No 3 above) and when balanced tubes have been
          loaded into the buckets of the rotor correctly as per point No 4 above, the lid of the centrifuge can be closed.

          To close the lid, place a small amount of pressure on the lid until the lid system locks (approx 6 seconds). A light on the lid
          button will come on when the lid has been closed properly.

          Setting Speed (RPM) / RCF
          Selection of the mode required (RPM or rcf)
          The button marked speed and rcf is used to select the speed / rcf required for the run. The light beside the particular mode
          selected will be lit to indicate which mode has been selected. To change the mode selected, press the button marked
          speed / rcf. Check that the light beside the mode you require is on.

          Selection of the right speed/rcf value
          Turn Knob 4 to the right and this will increase speed/rcf in increments of 500.
          Turn Knob 5 to the right and this will increase speed / rcf in increments of 10

          The preset value will be indicated on preset Speed Display
          The actual speed/rcf will be indicated on Actual Speed Display.



               Actual Speed         Select Speed          Preset Speed      Knob 4 Speed               Knob 5 Speed
               Display              Mode RPM or rcf       Display           Adjustments                Adjustments
                                                                            increments of 500          increments of 10




                                                                                                                           Lid




          The preset speed/rcf should not be higher than the maximum allowed speed/rcf for the rotor (7000 x g for the
          Swinging Bucket Rotor or 21,000 x g for the Fixed Angle Rotor).
          The maximum speed allowance depends on the rotor and the quality and volume of the tube:

          Selection of the operating time
          Turn Knob 3 to the right to go from 1 minute to 60 minutes. The time you set will appear in the digital display 1. For
          continuous runs turn knob 3 to the right to the limit stop. Continuous run logo ‘- -‘ will be indicated on digital display 1.
          Press the stop key to stop a continuous run.

          Adjustment of the radius correction factor
          The radius factor needs to be checked prior to every run to make sure that the centrifuge runs to the correct RCF value for
          each run.
          If you are using the Fixed Angle rotor with no adaptors, the radius correction factor should be set to 0
          If you are using the Swinging bucket rotor with no adaptors, the radius correction factor should also be set to 0
          If you are using the Swinging bucket rotor PLUS Adaptors, then the radius correction factor must be adjusted according to
          the rotor and type of adaptors being used :

     Tube          Rotor Type                                 Tube / Insert Type                                 Radius
     Rack                                                                                                        Correction Factor
     714.003       Swinging Bucket Rotor 220.80 V02           250 ml Tube Insert                                 2.8 cm
     714.007       Swinging Bucket Rotor 220.80 V02           50 ml Tube Insert Conical Blue Top Tubes           2.8 cm
     714.010       Swinging Bucket Rotor 220.80 V02           15 ml Tube Insert Conical Blue Top Tubes           3.1 cm

          IMPORTANT: Tube racks for 50ml Blue Top Conical Tubes and tube racks for 15ml Blue Top Conical Tubes
          CANNOT be spun at the same time. For further information regarding the Radius Correction Factor, see section
          on Adjustment of the radius correction factor.
___________________________________________________________________________________________________________
                                                                 Page 3 of 5
Safe Work Procedure                                                                             Date Effective: 01/01/2007
Uncontrolled document when printed                                                        Current Version: 1.2, 15/08/2007
                                               Press button to
                                               select Time or        Knob 3
                                               Radius Mode           Adjustments to
                                                                     Time / Radius




                                                                                                                         Knob to
                                                                                                                         adjust
                                                                                                                         Brake
 Display 1                                                                                                               Intensity
 indicating Time
 or Radius

                                                                                                                           Start
                                                                                                                           Button


                                                                                                                           Stop
                                                                                                                           Button




                                                                                          Knob to adjust Temperature
                                                                                          Settings


         Selection of Brake Intensity
         The brake intensity can be adjusted from 0 to 9 by adjusting the knob adjacent to the Brake button. The higher the brake
         intensity, the faster the rotor decelerates at the end of a spin.

         Setting the Temperature of a Run
         This centrifuge is a refrigerated centrifuge and the lowest temperatures that can be reached at maximum speed are:

                                                 Lowest Temperature of Z 513 K
 Rotor                                        Maximum Speed                                  Temperature (+/- 1°C)
 Fixed Angle 220.70 V06                       5000 rpm                                       +2°C
 Swinging Bucket Rotor                        15000 rpm                                      +5°C
 220.80 V02

         Pre-cooling
         This feature is used to avoid considerable temperature fluctuations at the beginning of a run at low temperatures. The
         centrifuge and the rotor are cooled together before the samples are inserted. To pre-cool, follow the steps below:
              1. Insert the rotor and the buckets required and secure the rotor to the shaft.
              2. Set the desired sample temperature and close the centrifuge lid
              3. Press ‘Pre-cool’ button. The centrifuge will accelerate up to the most suitable pre-cool speed and the preset
                   temperature will be reached in a few minutes.
         Press Stop button to finish the precool function.

         Starting the Centrifuge
         Once the rotor has been secured, tubes balanced and correctly loaded and parameters for the particular spin have been
         set up, the lid must be closed to start the run. Once the green light adjacent to the Lid button is lit, the centrifuge can be
         started by pressing the start button.

         Stopping the centrifuge
         The centrifuge can be stopped at any stage during a run if necessary by pressing the stop button.


         Maintenance
             •   The rotor chamber, rotor and accessories must be kept clean
             •   Monthly lubrication with acid free Vaseline of the bolts of the swing out rotor is required
             •   If rotors, buckets or tube racks come in contact with corrosive liquids including acids, alkaline solutions, solutions
                 containing heavy metals, anhydrous / chlorinated solvents and salt solutions, the contaminated parts must be
                 cleaned thoroughly by using a neutral cleaning detergent (eg Cleantec HI Clean Manual Dishwashing & Neutral
                 Cleaner) in warm water with a cloth / paper towel
             •   To maintain good hygiene and to reduce the risk of corrosion, cleaning using a neutral cleaning detergent (eg
                 Cleantec HI Clean Manual Dishwashing & Neutral Cleaner) with a ph 6 – 8 is required
             •   After cleaning all parts must be dried thoroughly by hand or in an oven with temperature of <= 50°C
             •   Treat anodized aluminium parts with anti-corrosion oil to reduce the risk of corrosion
             •   Regularly remove condensation with a cloth from the rotor chamber




___________________________________________________________________________________________________________
                                                          Page 4 of 5
Safe Work Procedure                                                                   Date Effective: 01/01/2007
Uncontrolled document when printed                                               Current Version: 1.2, 15/08/2007
          Cleaning after Glass Breakage
          If breakage of tubes occurs during a high speed run, the centrifuge, rotor, rotor, buckets and the rotor chamber must be
          thoroughly cleaned and all broken particles must be removed immediately. If this is not done, remaining particles could
          scratch the protective coating of the rotor. If the rotor chamber has not been properly cleaned, the resulting fine black dust
          produced can cause significant damage to the centrifuge chamber, rotor, buckets and samples.

          Disinfection
          If infectious material is spilt in the centrifuge, the rotor, rotor chamber, buckets and tube racks should be disinfected with a
          neutral disinfection agent eg Trigene Advance 1:10 dilution or undiluted Viraclean. Leave the Trigene Advance in contact
          with the material for a minimum of five minutes then rinse with demineralised water and dry thoroughly.

          For disinfection using Viraclean, apply undiluted Viraclean to the area requiring disinfection and leave for 10minutes. Use
          disposable toweling to wipe up the Viraclean and repeat the process. A disinfectant spray should be used to thoroughly
          clean the chamber, rotor, buckets and tube racks.

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

Samples to be spun, MSDS for samples, appropriate centrifuge tubes that are compatible for the chemicals / reagents being spun,
centrifuge, rotor, adaptors, hermetically sealed lid (for infectious samples), Lab coat, Gloves, closed shoes, Balance to 1 decimal
place which can weigh up to 1Kg, Neutral Cleaner detergent / disinfectant eg Viraclean, Trigene Advance


List potential hazards and risk controls including
specific precautions required

Hazard: the type & characteristics of the reagent to be spun
Risk Controls: Be familiar with MSDS of reagents and wear PPE according to the MSDS. As per Manufactures Manual, if any
contamination / spillage occurs, use an appropriate cleaning or disinfection procedure for the type of contamination. Never spin
corrosive or explosive materials in this machine.

Hazard: imbalance of the samples will cause centrifuge to vibrate / move and possibly come off the bench if imbalance is severe and
the set speed is high
Risk Control: Only trained personnel to use the machine – it is critical that all users understand how to balance the samples and how
to insert adaptors into buckets so that they are in the right position to be balanced.

Hazard: height of the centrifuge
Risk Control: Use safety step to access machine for inserting / removing samples, changing rotors.

Hazard: Weight / Size of the swing out rotor
Risk Control: Manual Handling Training required

List emergency shutdown instructions
This centrifuge has a number of inbuilt safety features:
     1. Imbalance: If loading is unequal (which will result in imbalance), then the operation will be interrupted in the acceleration
           stage and the rotor will be decelerated to standstill. An error message will be displayed on the unit.
     2. Over temperature: If the temperature reaches over 50°C, the rotor will decelerate and stop. Open the chamber to allow it
           to cool down and then restart the machine. If this occurs a number of times, contact In Vitro Technologies Service Team
     3. In the case of power failure, or any malfunction, the lid can be opened manually at the side of the machine by following
           instructions in the Manufacturers Instruction Manual
     4. There is an emergency manual lid release which can be used to retrieve samples in the case of a power failure. Once the
           centrifuge has been switched off and the power plug has been unplugged, remove the plastic plug on the left side of the
           centrifuge housing. Use the rotor tool to loosen

List clean up and waste disposal requirements
To clean the centrifuge interior a neutral detergent (eg Cleantec HI Clean Manual Dishwashing & Neutral Cleaner) should be used to
pro long the life of the machine and to prevent corrosion.


List legislation, standards and codes of practice used
in the development of the SWP
Instruction Manual for Bench Top Centrifuge Z 513 K



Supervisory approval, training, and review
Supervisor: Prof Beng Chong                                           Signature:

Plant custodian: Prof Beng Chong                                      Signature

List competency required – qualifications, certificates, licencing, training - eg course or instruction: Training in this SWP and Manual
Handling Training is required.


SWP review date: September 2010          Responsibility for SWP review: Sinead O’Reilly

___________________________________________________________________________________________________________
                                                          Page 5 of 5
Safe Work Procedure                                                                   Date Effective: 01/01/2007
Uncontrolled document when printed                                               Current Version: 1.2, 15/08/2007

				
DOCUMENT INFO
Shared By:
Categories:
Stats:
views:45
posted:3/30/2010
language:English
pages:5