NIHR Sheffield Clinical Research Facility. Use of the Refrigerated Centrifuge

Transcription

1 * Standard Operating Procedure NIHR Sheffield Clinical Research Facility Use of the Refrigerated Centrifuge SOP History (archived date) V1.2 (01/11/2010) reviewed by John Humphreys, V1.1 (01/11/2010); V1.0 (03/03/2010) SOP Number CRF.L109 Created by Emma Goodwin Version 1.3 Date Review Date October 2013 Related SOPs 1 CRF.L105 Approved by Alison Jenkins Page 1 of 6 *The master document is available on the CRF I:\ drive and website: print-off of this document by anyone other than the CRF SOP Co-ordinator will be classed as uncontrolled. Researchers and their teams may print off this document for training and reference purposes but are responsible for regularly checking with the CRF Sop Co-ordinator for the most recent version.

2 Standard Operating Procedure: NIHR Sheffield Clinical Research Facility This SOP has been written to give general guidance to study personnel on how to use a centrifuge. There are no Good Laboratory Practice Guidelines concerning the use of a centrifuge. Background It is important that all staff working in the Clinical Research Facility use the same procedure when operating the centrifuge as specified in the trial protocol, to ensure consistency and continuity in the recording of results. Centrifuges processing pathological material can create considerable health risks by liquid spillage and droplet dispersion. It is important that they are properly designed and constructed, installed, operated and maintained. They should be sited so that operators can see into the bowl and can easily load the buckets and containers. The LABOFUGE 400 R, Heraeus Megafuge 1.0 R and Heraeus 16 R are designed to perform a variety of separation tasks in small-scale laboratories. All units are constructed as bench-top models. Definition Centrifugation is the process of separating two or more liquids in a mixture by rotation in a container so that the lighter density liquid rises to the top. It is occasionally necessary to centrifuge samples in refrigerated conditions to ensure sample integrity. Procedure 1. The investigator is responsible for ensuring that the centrifuge is used according to the manufacturer s instructions and according to the study protocol. This duty can be delegated to other appropriately qualified members of the research team as recorded on the project delegation log. Manuals for each centrifuge device can be found in the laboratory in the Clinical Research Facility. 2. The investigator or delegated person will refer to the protocol to ensure specific requirements for the use of the centrifuge are identified. 3. When working in the laboratory the investigator or delegated person must wear protective clothing. This may be in the form of a uniform, a laboratory coat or a disposable apron. Laboratory coats are available in the Clinical Research Facility laboratory. Aprons can be found laboratories and clinical areas of the CRF. 4. The investigator or delegated person should wash their hands before handling blood samples according to STH hand hygiene policy. 5. The investigator or delegated person should wear disposable gloves when there is a risk of contamination. A supply of disposable gloves in various sizes is available in the Clinical Research Facility laboratory. 6. The investigator or delegated person must ensure that the centrifuge is switched on at the wall plug socket. 7. The lid of the centrifuge will be open when it is not in use. The investigator or delegated person must open the lid fully so the inside of the centrifuge can be seen. Page 2 of 6

3 8. The investigator or delegated person must unscrew the lid of the rotator bucket to be used. 9. The investigator or delegated person must place the blood tubes in the correct swinging bucket adaptors located inside the rotator buckets. 10. The investigator or delegated person must ensure that the centrifuge is balanced, making sure on opposite sides there is an identical tube balanced by weight. Spare balance tubes can be found in a rack next to the centrifuge in the Clinical Research Facility laboratory. Please refer to the manufacturers instructions for further information on balancing instructions, these can be found in the Clinical Research Facility laboratories. 11. The investigator or delegated person must ensure that the rotator buckets are firmly closed with the covers in place for each centrifugal run. 12. The investigator or delegated person will set the time on the centrifuge according to study protocol. 13. The investigator or delegated person must set the time speed using the set key on the time/rotor field on the front panel of the centrifuge and scroll up or down using the + and keys until the desired time is reached. Press set a second time on the time/rotator field and this will programme the time. The set time will be displayed in red on the front panel of the centrifuge. 14. The investigator or delegated person must set the speed on the centrifuge according to study protocol. The LABOFUGE 400 R, Heraeus Megafuge 1.0 R and Heraeus Megafuge 16 R has speed set automatically in revolutions per minute (RPM). 15. The investigator or delegated person can covert RPM if this is stipulated in the study protocol to relative centrifugal force (RCF) by pressing and holding in the set button. When this is pressed rc will show in red on the time rotator display. The display panel will flash with numbers in red. The investigator or delegated person must scroll up or down pressing the + and - keys until the desired speed is reached. 16. The investigator or delegated person must note that when samples are spinning the display panel will show in RPM. If the set button is held in the speed will display in RCF s. 17. The investigator or delegated person must set the speed rcf. The speed will be shown in the speed/rcf field and this can be changed using the speed/rcf button, the speed can be changed by scrolling up or down using the + and keys until the desired speed is reached. Press set a second time and this will programme the speed. 18. When set, the speed will revert to zero and build to the set speed gradually. This will allow for any problems to be detected and resolved. The centrifuge can be stopped in an emergency by pushing the stop/pause button and waiting for the device to slow to a stop and release the door lock (please refer to Appendices Glass Breakage and Blood Spillages in the Centrifuge). 19. The LABOFUGE 400R, Megafuge 1.0 R and 16R is a refrigerated centrifuge. Room temperature is set at 20 O C on the centrifuge. Temperature is displayed in red on the front panel of the centrifuge. Page 3 of 6

4 20. If refrigerated centrifuge samples are required the investigator or delegated person must set the temperature at least 10 minutes before use and the lid must be closed. 21. The investigator or delegated person must set the temperature of the centrifuge according to study protocol. 22. The investigator or delegated person can set the temperature by pressing the set keypad of the temp field. Temperature value can be changed in O C. Press set a second time and this will programme the temperature. 23. The investigator or delegated person should be aware that items will be cold when removed from the refrigerated centrifuge unit. 24. After use the investigator or delegated person must reset the temperature at 20 O C and leave the lid of the centrifuge open to avoid condensation. 25. The investigator or delegated person must never leave the centrifuge in refrigerated mode with the machine switched on and the lid open. This will cause the refrigeration mechanism to break. 26. To start the centrifuge press the start/stop key on the front panel of the centrifuge. After the centrifuge has started the lid key will become disabled and the red light will be unlit. 27. Once the centrifugal run is complete the investigator or delegated person must open the lid of the centrifuge by pressing lid key on the on the front panel of the centrifuge, when there is a yellow light this indicates that the lid can be opened. After the lid is opened, OPEN will be indicated in the speed/rcf display. 28. If the buckets are stuck in a horizontal position then the investigator or delegated person must try to push them to their vertical position. If unable to do so please see the manufacturer s instruction manual for further instructions. The manual can be found in the laboratory in the Clinical Research Facility. If problems still persist the investigator or delegated person must inform a member of the Clinical Research Facility team who will call the manufacturer for their assistance. 29. The investigator or delegated person must then remove the rotator bucket from the centrifuge ensuring that it is kept upright in a vertical position and not tipped up. 30. The investigator or delegated person must unscrew the lid of the rotator bucket, ensuring that the tubes are not tipped. 31. The investigator or delegated person must remove all blood tubes from the swinging buckets and place in a storage rack next to the centrifuge. 32. Once the blood has been placed in the storage racks the investigator or delegated person must replace all buckets and lids and turn off the machine at the socket. 33. The investigator or delegated person should remove gloves, coats and gowns before leaving the laboratory. Coats and gowns should be left close to the exit. Laboratory coats should not be placed in personal lockers. Page 4 of 6

5 34. The investigator or delegated person should wash their hands after handling blood samples according to STH hand hygiene policy. Related Documentation 1 Document Name Safe working and the prevention of infection in clinical laboratories and similar facilities LABOFUGE Operating Instructions STH Hand Hygiene Policy Project Delegation Log Sheffield Infection Control Guidelines Author Butterworths Online Direct Service Heraeus STH Infection Control Team CRF STH Infection Control Team 1 The location(s) of any related document(s) are listed in the CRF SOP Referenced Documents Directory. The CRF SOP Referenced Document Directory and any Related SOPs, listed on page 1 of this SOP, can be accessed electronically at or can be requested by contacting Sheffield Clinical Research Facility, O Floor, Royal Hallamshire Hospital, Glossop Road, Sheffield, S10 2JF, Appendix When using the centrifuge the following points should be adhered to: Sealed buckets or rotators must be used. Check that bucket seals are intact so that they provide adequate protection against liquid dispersion in the event of an accident during use. The containers being centrifuged must be strong enough to withstand the centrifugal forces to which they will be exposed. The bucket sealing rings must be inspected regularly and changed as necessary. The fluid being centrifuged should be introduced into the container carefully, the threads or outside of the container must not be contaminated. Containers must be filled according to the marker s instructions. At least 2cm of headspace must be left between the liquid level and the container rim. Excessive pressures can be generated in overfilled containers which may also lead to leakage. On a weekly basis or after 20 runs the inside of the centrifuge and its buckets should be cleaned decontaminated and greased. Cleaning products can be found in the laboratory in the Clinical Research Facility. Rotator and bowls should not be soiled and can be wiped down with Trigene and alcohol they should then be thoroughly dried with a dry cloth. Buckets and inserts that are not obviously soiled can be treated in the same way but any soiled components should be decontaminated appropriately. Greasing the buckets ensures smooth movement and less vibration during acceleration and deceleration of the motor. Please see manufacturer s instructions which can be found in the Clinical Research Facility laboratory. Page 5 of 6

6 Glass Breakage CONTROLLED DOCUMENT- DO NOT COPY Centrifuge glassware shows a correspondingly higher failure rate with rising g-forces. If the investigator or delegated person does experience a glass breakage, they should inform a senior member of the Clinical Research Facility team. If a breakage occurs in the centrifuge the lid should remain closed for 30 minutes to allow for any glass or soluble particles to settle. Once the lid is open the glass splinters must be removed immediately and completely from the buckets and rotator chamber. The glass splinters can damage the surface protection or could get stuck on the trunnions of the swing out rotators where they can impair smooth movement of the swinging buckets. When glass splinters get into the rotor chamber and are whirled around by air circulation, a very fine (black) metal dust can develop. Metal abrasion can contaminate the rotor chamber, rotor buckets and samples. With a strip of Vaseline or trunnion grease applied vertically on the chamber wall (from top to bottom), the glass dust can be removed with a smooth centrifuge cycle. Please refer to the manufacturer s instructions that can be found in the CRF laboratory. Blood Spillages in the centrifuge If the investigator or delegated person does have a blood spillage whilst using the centrifuge, they should inform a senior member of the Clinical Research Facility team. Any liquid spilt in the centrifuge should be cleaned and decontaminated according to the manufacturers instructions. Manufacturer s instructions can be found in the Clinical Research Facility laboratory. Please refer to SOP CRF.L105 Procedure for dealing with spillages and Sheffield Infection Control Guidelines. Page 6 of 6