Centrifuge STANDARD OPERATING PROCEDURE

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STANDARD OPERATING PROCEDURE Centrifuge Model: Sorvall, RC-5B Plus Manufacturer: Kendro Laboratory Products Location: Technology Transfer Lab, 1568 Food Sciences Building Publication Date: 09/26/2014 Iowa State University does not discriminate on the basis of race, color, age, ethnicity, religion, national origin, pregnancy, sexual orientation, gender identity, genetic information, sex, marital status, disability, or status as a U.S. veteran. Inquiries regarding non-discrimination policies may be directed to Office of Equal Opportunity, 3350 Beardshear Hall, 515 Morrill Road, Ames, Iowa 50011, Tel. 515 294-7612, email eooffice@iastate.edu.

Description and Uses The Sorvall Model RC-5B Plus is a high-speed, microprocessor-controlled refrigerated floor-model centrifuge used to separate substances of different sizes or densities. The centrifuge operates over a temperature range of -20 C to +40 C and a time range of up to 120 min (plus a Hold setting that allows operation without a time limit). The centrifuge has a speed selection range of 500-21,000 (± 100) rpm and a maximum relative centrifugal force of 51,427 x g (using the F-20/Micro Rotor). Power Specifications Motor: Spindle driven, Sorvall RC-5B+, SN 9801276 Voltage/Amperage: 200-240V/Single Phase/30A Speed Settings: 500-21,000 (± 100) rpm @ 60 Hz. Temperature Range: -20 C to +40 C Time Settings: 0-120 minutes or HOLD For further information on specifications, general operation and installation requirements, see Chapters 1 and 2 in the Sorvall RC-5B Plus Operating Instructions manual (filed in the drawer closest to the centrifuge in the Technology Transfer Lab, 1568 Food Sciences Building). Potential Hazards and Safety Precautions Electric Shock/High Voltage (200V-240V) Make sure that the wall outlet receptacle is properly wired and grounded, and matches the instrument s power cord and plug. Make sure the area around the outlet, floor and your hands are completely dry when plugging or unplugging the electrical cord to/from the outlet. Fire Safety/Explosion Hazard Do not use the centrifuge with materials capable of developing flammable or explosive vapors. Note: No flammable liquids are to be centrifuged! Spindle-driven Pinch Points/Possible Entanglement of Extremities, Hair, Jewelry or Clothing Use only Sorvall rotors and accessories designed for this centrifuge. Do not exceed maximum speeds for the rotors used. Never try to slow or stop a rotor by hand. Make certain the chamber door is locked and secure before operating the centrifuge. Never try to override the door-interlock system while the rotor is spinning. Make sure to secure long hair and any loose clothing or jewelry before operating the centrifuge. 2

Biohazard/Possible Illness or Physical Damage to Tissues and Organs Normal operation may involve use of pathogenic, toxic, radioactive, or otherwise hazardous materials. Such materials should not be used in the centrifuge unless all necessary safety precautions have been taken (see Operating Instructions manual). Note: Never use flammable solvents in a centrifuge! Be sure to wear all required personal protective equipment that is listed below. Required Personal Protective Equipment Lab Coat Hair Net (Tie Back Long Hair) Safety Glasses or Goggles Rubber, Latex or Disposable Gloves No Loose Fitting Clothing No Open-toed or Open-heeled Shoes Training Required Training *Denotes courses offered online Machine & Site-Specific Training Fire Safety & Extinguisher Training* Laboratory Safety: Core Concepts* Recommended Training for Frequent Users Electrical Safety & Lockout/Tagout Laboratory Safety: Spill Procedures 3

Operation 1. Be sure to acquire all required training, including site-specific training before operating the centrifuge. 2. The Operating Instructions manual (Chapter 3) and the Rotor Instructions manual provide a more complete description of how to operate the centrifuge and its rotors. Be sure to read these manuals before using the centrifuge to become familiar with its correct operation and individual component parts. Both manuals are on file in the Technology Transfer Lab, 1568 Food Sciences Building. 3. Be sure to wear all required personal protective equipment. 4. Press the power switch to the ON position. The power switch is located in the upper right corner of the front cabinet panel, just below the chamber door button (see Figure 1). 5. To open the chamber door, push the DOOR button. This is a large rectangular button labelled DOOR and is located in the upper right corner of the front cabinet panel, just above the power switch (see Figure 1). Chamber Door Door Button Figure 1 Door Handle Power Switch Control Panel 6. The rotor that is supplied with this centrifuge is the Sorvall SLA-3000 fixed-angle rotor, which has a capacity for six 500-ml centrifuge bottles (see Figure 2). The maximum speed for the Sorvall SLA-3000 fixed-angle rotor is 12,000 rpm (maximum RCF = 24,336 x g). Other rotors for this centrifuge can be ordered through Kendro Laboratory Products (see contact information provided on page 1 of this Standard Operating Procedure). Note: Never exceed the maximum RPM or RCF specifications listed for any rotor, as serious damage to the centrifuge will result. If you plan to centrifuge your sample under refrigeration, it may be best to pre-cool the rotor either in the centrifuge chamber or in a refrigerator/cold room before operation. Please make note that our rotor is RED, not black as shown in Figure 2. Figure 2 4

7. Remove the lid and safely install Sorvall SLA-3000 fixed-angle rotor (or another ordered Sorvall rotor) in the chamber according to the directions given in the Rotor Instructions manual. Note: Be sure that the rotor selected is compatible with the centrifuge. Rotors used MUST HAVE drive holes in which the spindle s drive pins fit into. DO NOT USE ROTORS WITHOUT COMPATIBLE DRIVE HOLES. To check for drive holes, invert the rotor and look in the center hole. When installing the rotor, be sure that that drive holes are properly seated on the drive pins of the hub (spindle). Also, make sure that the rotor center hole and tapered spindle are clean and free of nicks or scratches. Wipe surfaces clean before each operation to reduce the chance of the rotor sticking to the spindle. 8. Fill the 500-ml centrifuge bottles with samples to be centrifuged. Note: Equally balance the bottles in pairs, otherwise the rotor will vibrate, disturb and resuspend the solids and could potentially damage the centrifuge. Once balanced, insert each bottle of the balanced pair opposite one another in the rotor. 9. Replace the lid and tighten it onto the rotor as per the directions given in the Rotor Instructions manual (on file in the drawer closest to the centrifuge in the Technology Transfer Laboratory, 1568 Food Sciences Building). After making sure the rotor and lid are secure, carefully close the chamber door. 10. For the following instructions, refer to the Control Panel schematic in Figure 3. a. Using the Blue & Red TEMPERATURE control knobs, set the desired temperature range. b. Turn the TIME dial to the desired run time or set to HOLD to allow operation without a time limit. c. Turn the SPEED dial to the desired RPM setting. Note: For the Sorvall SLA-3000 fixed-angle rotor, the maximum allowable speed is 12,000 rpm (maximum RCF = 24,336 x g). d. If the braking function is needed to slow the rotor after a spin, turn the brake ON by pressing the BRAKE switch until the indicator light comes on. Temperature Control and Meter Speed Meter and Dial Time Dial Brake Switch and Light Door Light Start Button Imbalance Light Figure 3 11. Once the TEMPERTURE C indicator lies within the preset temperature range, press the ST=ART button. Then, the centrifuge will accelerate to the selected speed and run for the selected time, unless an imbalance is indicated (see step 12). 5

12. If the IMBALANCE light comes on, then a bottle imbalance is indicated and the centrifuge will not start. If this is the case, rebalance your bottles, reinsert them properly in the rotor, secure the rotor and lid, close the chamber door, and try to START the centrifuge once again (refer to Figure 3). 13. Once spinning, allow the centrifuge to run for its pre-selected time and speed. 14. When the selected time expires, the rotor will coast or brake to a stop. At this point, the DOOR light turns on to indicate that the latch interlock has released (refer to Figure 3). Once released, open the chamber door by pushing the DOOR button and lifting up on the door handle (refer to Figure 1). Note: NEVER try to bypass the chamber door s interlock system. 15. Loosen the lid from the rotor and remove it. Then, retrieve the centrifuged samples from the rotor. 16. Repeat the listed procedure if more centrifugation is needed. When all samples have been centrifuged, press the power switch to the OFF position (refer to Figure 1). Record your use in the available centrifuge log book located in the drawer nearest the centrifuge. 17. Initiate clean-up procedures. Clean-up Procedures 1. Wash all centrifuge bottles with warm soapy water. 2. Rinse the centrifuge bottles with hot tap water, followed by a cool, distilled-water rinse. 3. If necessary, remove the rotor and wash/rinse as listed in steps 1 and 2. 4. Allow the clean bottles and rotor to dry by placing them into a drying cart or placing them upside down on a pad of clean paper towels. 5. If there was any spillage in or on the centrifuge, be sure to wipe all surfaces clean with an absorbent Teri towel. Then, wipe with a damp, but clean, Teri towel to rinse the surfaces. 6. Clean the surrounding work area. 7. Before leaving the work area, initiate check-out procedures by having the centrifuge and rotor inspected by the lab manager. Machine Care and Maintenance Report all operational problems to the laboratory or pilot plant manager. Centrifuge must be properly cleaned and inspected after each use. All final inspections are performed by the laboratory or pilot plant manager. The spindle base should be properly lubricated on a regular basis by the laboratory manager, pilot plant manager, or a trained service technician. 6

Accessories This centrifuge comes equipped with the Sorvall SLA-3000 fixed-angle rotor. Note: For this rotor, a maximum of 12,000 RPM (RCF = 24,336 x g) must not be breached. Additional rotors with their own set of specifications are available for purchase through the manufacturer. Condensed Operating Instructions manual (located in the drawer nearest the centrifuge, Technology Transfer Laboratory, 1568 Food Sciences Building). Condensed Rotor Instructions manual (located in the drawer nearest the centrifuge, Technology Transfer Laboratory, 1568 Food Sciences Building). Wrench (9/16 inch) Motor Brush Replacement Kit 7