PROPER USE OF THE SORVALL WX 80+

ULTRACENTRIFUGE

SOP # 01-REP

Staff Members, PIs and Students are responsible for following

Responsibility: this SOP. The designated lab supervisor is responsible for
ensuring that this SOP is followed.

This SOP covers the proper and safe use of the Sorvall WX+
Scope: Ultracentrifuge. It is applicable to all staff members/students
working with this machine.

Review Date: 06/04/2019

Ultracentrifuge: A device in which materials of different

densities are separated by rotation at various speeds about a
fixed, central point, by way of centrifugal force. The denser
particles tend to move along the length of the tube to a greater
Definitions radius of rotation, displacing the lighter particles to the other
end
Rotor: The rotating part of the centrifuge, which holds the
samples.

1. PL-ES-006-RA-xxxx - Using a Centrifuge

Related
2. Sorvall WX Ultracentrifuge - User's Manual
Documents:
3. Sorvall WX Ultracentrifuge - Inspection, Maintenance and
Service Record
4. Sorvall WX Ultracentrifuge and Rotor Log Sheet
5. Sorvall WX Ultracentrifuge - Documentation of Training

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 - SOP Outline

Procedures
Summary of Significant Health and Physical
3
Hazards
Personal Protective Equipment Required 3
Risk Assessment 3
Emergency Contacts 3
Preparatory Steps 3
Warnings 3
Laboratory Procedures
(1) Preparation 4
(2) Centrifuge Operation 5
(3) Emergency Procedures 9
(4) Maintenance 9
(5) Service 9
(6) Cleaning Procedures 10
(7) Training 10
References 11
Procedures Quality Assurance 11

Appendix
I. Equipment Details 12
II. List of Compatible Rotors 13
III. Ultracentrifuge and Rotor Log Sheet 14
IV. Acceleration and Deceleration Modes 15
V. Operation Flowchart 16
VI. Troubleshooting Table 17
VII. Inspection, Maintenance and Service
20
Record
VIII. Documentation of Training 21

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- Summary of Significant Health and Physical Hazards

Physical Hazards: Mechanical stress, metal fatigue and

corrosion can lead to mechanical failure over time. Unbalanced
centrifugation and other mechanical failures can result in
centrifuge explosion.

Exposure Hazards: Centrifuged materials can become

aerosolized, exposing the user to hazardous air when the
centrifuge is opened.

- Personal Protective Equipment Required

(1) Protective Gloves
(2) Lab Coats
(3) Closed-toe Shoes
(4) Safety Eye Goggles

- Risk Assessment
-
The overall health and safety risk of this procedure is described
in Ref: PL-ES-006-RA-xxxx - Using a Centrifuge

- Emergency Contacts
-
List Of QU Emergency Contacts Here

- Preparatory Steps

This ultracentrifuge must not be used without the approval of the

equipment custodian (Mr. Saleh, Tel: 7837, Room C-104).

(1) Review the emergency information, special handling and waste

disposal procedures contained in this SOP.
(2) Visually verify that the access to the emergency
eyewash/shower unit and fire extinguisher are not blocked.
(3) Visually verify that access to the emergency doors are free of
obstructions
(4) Use the appropriate protective equipment
(5) Make an entry in the device logbook before each use.

- Warnings

(1) Make sure that the rotor is appropriate for the instrument and
use the lid that matches the rotor. Rotor can fail due to
excessive speed, causing catastrophic failure of centrifuge.

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 Know the speed limits of the rotor and do not exceed them
(See Appendix II).
(2) Rotor imbalance can cause catastrophic failure of centrifuge
explosion), injuring user.
(3) Swing bucket rotor are designed to be used with all the buckets
in place, even if some of them are empty. Do not use this type
of rotors without all the buckets in place.
(4) Rotor can fail due to extensive wear and tear, causing
catastrophic failure of centrifuge or even injuring the user.
Inspect all the rotors regularly and replace if damaged.
(5) Centrifuge tubes can break inside the rotor due to excessive
wear, tear or inappropriate use causing catastrophic failure.
Ensure that you use the correct tubes for your samples, and do
not use cracked or broken tubes. Phenol and chloroform will
rapidly destroy polycarbonate tubes and must be used in
polypropylene tubes.
(6) Plastic centrifuge tubes not certified as reusable from the
manufacturer, should be discarded after one cycle of
ultracentrifugation. The failure rate for used tubes is a hazard
that justifies using new tubes for each high G run.
(7) Do not lean or place items on the instrument while it is
operating.
(8) Do not leave the centrifuge until full operating speed is
attained, and the instrument appears to be running normally
without vibration.
(9) If vibration occurs, stop the run immediately; wait until the
rotor stops, and check the load balances.
(10) In event of a power failure, do not try to open the lid to retrieve
samples for at least one hour.

- Laboratory Procedures

(1) Preparation

- Prepare suspensions in appropriate tubes. Reusable

ultracentrifuge tubes have a maximum volume and a minimum
volume, and the maximum speed of the tube depends on the
volume in the tube. The maximum volume level in each tube
should not exceed 75% of the tube capacity.

- Make sure that the sample tubes are appropriate for your
samples and the speed required. All tubes used should be of the
same type. Check all tubes for signs of stress and/or crack.
Discard any tubes if necessary.

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 (2) Centrifuge Operation

Figure 1: External View of

Ultracentrifuge

- Turn on the POWER switch. Wait for the initial screens to

appear on the touchscreen. The chamber door will unlock.
- Open the chamber door and make sure that the chamber is dry.
If there is moisture present, wipe it with a clean dry cloth.
- Open the door completely, gently install the rotor on the crown
and close the door. Ensure that the rotor is firmly attached to
the spindle and that the rotor lid is properly secured before
starting the machine.

Figure 2: Rotor installation procedure

- Always balance the centrifuge tubes (by weight, not just by

eye), and put the balanced tubes in symmetrical positions in the
rotor. Balance all samples to be within 0.05 gr difference. The
rotor’s center of mass should always be on the rotation axis. If
necessary, use water samples to balance rotor out.

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 Figure 3:
Symmetrical
Placement of Tubes
on a 12-Position
Rotor

- Set the Speed, Time and Temperature parameters of the run on

the display screen.

Figure 4: Initial Screen and Run Screen

- Press the SPEED, TIME, TEMP, ACCEL or DECEL field on

the touchscreen to display the on-screen keypad.

- Press the field of the item you want to

set. The initial digits are displayed in
blue.

- Press the on-screen keypad to

enter a numeric value. Example:
For 100,000 rpm press [1] [0] [0]
[0] and then [Enter]. If you enter
the wrong value, press the [CE]
button and enter the correct value. Figure 5: Setting Speed Parameters

- When more than one SPEED, TIME, or TEMP run parameter is

selected, you do not need to press the [Enter] button. When you
touch the field of another run parameter, the currently entered
item is confirmed.

- To set continuous run (HOLD) and specify the time during

operation, enter the sum of time elapsed so far plus the desired

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 remaining run time. For example, if the centrifuge has been
running for 5 hours and you want operation to stop 1 hour and
30 minutes later, touch the TIME field and enter [6][ : / . ] [3]
[0].
Acceptable Values
Speed: Set a value between 1,000 rpm to maximum speed in
100 rpm increment
Time: Set a value up to 999 hours and 59 minutes in 1-minute
increments.
Temp: Temperatures in the 0 to 40°C range can be set in 0.1 ℃
increments.

- To set the acceleration and

deceleration mode press a field to
display the on-screen keypad and
then press the field whose value you want to set. The numeric
values are displayed in blue. Press the on-screen keypad to enter
a numeric value and then the [Enter] button. If you enter the
wrong value, press the [CE] button and enter the correct value.

Acceptable Values (See Appendix IV for more information)

ACCEL: 1 to 9, 0
DECEL: 1 to 9, 0 or Free coast (FREE)

- To select a rotor, press the Rotor

indicator field in the Run screen.

- Check that the desired rotor is displayed. If it is, select it and

press the [OK] button. When the selected rotor field turns
blue, press the [OK] button to return to the Run screen.

- When the Rotor management screen does not display the desired
rotor, press the [Rotor Catalog] button. In the ROTOR
CATALOG screen, select the desired rotor type. In the screen
displaying rotor types, press the field of the desired rotor. Check
that the pressed field turns blue and press the [Enter] button.

- To confirm rotor details, press the rotor field to turn it blue and
press the [Details] button. A window that allows you to check
rotor information appears. If this is the selected rotor, press the
[Enter] button. To select a different rotor, press the [X] button
to return to the Select rotor screen.

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 Figure 6: Rotor Selection
Screen

- The Run screen

appears and the
name of the
selected rotor
type is displayed
in the Rotor
indicator field

Figure 7: Screen display after a successful rotor

selection

- Press the [VACUUM] button if necessary (You

can skip this step). The indicator of the
[VACUUM] button displays the vacuum level in
the rotor chamber. Set a high vacuum level before pressing the
START button to process samples sensitive to temperature
changes.

- Press the [START] button. The [START] button

lamp then starts to blink and the rotor is now turning.
The timer begins operating. If the actual run timer is
set, the timer will start operating when set speed is reached.
When the set speed is reached, the [START] button lamp
changes to steady lighting and a white dot of light starts to spin
around the button.

- The operation stops when the set centrifugation time

has elapsed (end of run) or when the [STOP] button
is pressed.

- When the rotor stops fully, the [STOP] button changes to

steady lighting and the buzzer sounds to indicate the rotor has
stopped.

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- Press the [VACUUM] button. The vacuum pump stops, the air
leak valve starts operating to return the rotor chamber to
normal atmospheric pressure. The door lock is released and the
door can be opened. When the rotor chamber returns to normal
atmospheric pressure, the "Now open the door" message
appears above the [VACUUM] button.

- Gently remove the rotor when it has stopped.

(3) Emergency Procedures

In the event that an incident or accident related to

centrifugation occurs:
(a) Turn off centrifuge and disconnect it from the power
source.
(b) Notify others in laboratory and evacuate.
(c) Notify the lab supervisor and the HSSE office.
(d) Do not use the machine again until it has been inspected
and repaired by a qualified service technician. Post warning
signs so no one plugs the machine back in.

(4) Maintenance (See Appendix VII)

- Check rotor and rotor bores regularly for deposits or damage.

- Some rotors come with an expiration date marked on the rotor
or rotor accessories. Rotors and rotor accessories must not be
used past their expiration date.
- Record warranty periods and retirement recommendations for
each class of rotor. Retiring rotors when the warranty expires is
prudent.
- O-rings and seals on rotors and safety cups should be inspected
routinely for breakage or damage and replaced promptly to
ensure effective containment.
- In case repair is needed, the ultracentrifuge should be tagged
and locked-out while awaiting service (see SOP-01:
Lockout/Tagout).
- All maintenance records should be gathered and kept in the
lab.

(5) Service (See Appendix VII)

Service Requirements: Annual inspection and service by

approved service agent.

Records: All service records should be gathered and kept in the

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 lab.

(6) Cleaning Procedures

(a) General Cleaning

- Use neutral agents for cleaning purposes and use alcohol-based

disinfectants for disinfection purposes. Do not use acetone,
caustic detergents, or detergents that contain chlorite ions.
- Remove any residues present in the bores of the rotor using a
bottlebrush and a hand-hot cleaning solution. Rinse well. Clean
bores with a dry cloth and allow them to dry.
- Allow rotor to accelerate once to maximum speed after
cleaning
- Autoclave the rotor in the event of contamination caused by
biohazardous substances. Never autoclave the rotor with the
lid attached. You should not use UV, beta, gamma, or any
other high-energy radiation source for disinfection. Do not use
any gassing for disinfection.

(b) Cleaning Spills

- Depending on the nature of the spill, clean up as described in

SOPs for Biohazard Spills, Flammables, Corrosives, or Toxic
Substances.
- If a breakage and /or spillage occurs, clean rotors and
centrifuge chamber immediately with 80% ethanol. The rubber
sealing may deteriorate if spills are not attended to promptly
and properly.
- Any large spills of hazardous materials or incidents resulting
in injury must be reported to the lab supervisor immediately.

(c) Cleaning of reusable centrifuge tubes and bottles

To clean centrifuge tubes and bottles:

- Wash with mild detergent in warm water, rinse them
thoroughly with clean water and allow them to air dry.
- Avoid cleaning plastic containers in automated dishwashers or
glassware washes because they are frequently too hot.
- Autoclave tubes only if necessary, keep the temperature at or
below 100ºC and autoclave for 15 minutes (Autoclaving may
void the warranty for the container. Check with the
manufacturer to make sure autoclaving is safe.).

(7) Training (See Appendix VIII)

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 Users of this device must have attended an instrument-specific
training session with the equipment custodian or the facility
approved lab manager prior to any use, covering general use
and safe practices of the instrument in question.

References

(a) Sorvall WX Ultracentrifuge - User's Manual

Quality Assurance Improper centrifugation speed, temperature and time can have
significant effects on sample quality. To ensure the validity
and reproducibility of the centrifugation protocols used, the
machine’s speed of rotation, chronometer and thermometer
must be verified once a year, and if necessary, calibrated by
authorized technicians. All calibration certificates should be
gathered and kept in the lab.

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APPENDIX

I. Equipment Details:

Model: SORVALL WX 80+

Manufacturing Year: 2015
Serial Number: 41887026
QU ID Tag: 0236000000000000000023F7
Max. Kinetic Energy: 1080 kNm
Voltage: 240V
Amperage: 16A
Power: 4800W
Location: H10 - Zone 4 - C-107 (Common Lab II)
Alaaedin Saleh
Custodian: Tel: +974 4403 7837
Email: alaaeldin.saleh@qu.edu.qa

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II. Compatible Rotors (Non-exhaustive list)

SORVALL WX 80+ - Compatible Rotors (Thermo Scientific)

Rotor
Model Maximum RPM Supported Tubes
Type

BIOFlex HC Rotor Swinging-bucket rotor 5.500 4 x 1000 mL

BIOFlex HS Rotor Swinging-bucket rotor 7.000 4 x 400 mL

TH13-6x50 Rotor Swinging-bucket rotor 13.100 6 x 50 mL

Fiberlite F9-6x1000 LEX Fixed-angle rotor 9.000 6 x 1000 mL

Fiberlite F10-4x1000 LE Fixed-angle rotor 10.500 4 x 1000 mL

Fiberlite F12-6x500 LEX Fixed-angle rotor 12.000 6 x 500 mL

Fiberlite F14-6x250 Fixed-angle rotor 14.000 6 x 250 mL

Fiberlite F14-14x50cy Fixed-angle rotor 14.000 14 x 50 mL

A21-24x15 Fixed-angle rotor 21.500 24 x 15 mL

Fiberlite F20-12x50 LEX Fixed-angle rotor 20.000 12 x 50 mL

Fiberlite F21-8x50y Fixed-angle rotor 20.000 8 x 50 mL

A22-24x16 Fixed-angle rotor 22.000 24 x 16 mL

A23-6x100 Fixed-angle rotor 23.500 6 x 100 mL

Fiberlite F23-48x1.5 Fixed-angle rotor 23.000 48 x 1.5 mL

A27-6x50 Fixed-angle rotor 27.000 6 x 50 mL

A27-8x50 Fixed-angle rotor 27.000 8 x 50 mL

T29-8x50 Fixed-angle rotor 29.000 8 x 50 mL

Fiberlite F50L-24x1.5 Fixed-angle rotor 50.000 24 x 1.5 mL

TCF-20 Continuous Flow Continuous flow rotor 20.000 1350 mL

TCF-20 Zonal Zonal rotor 20.000 1350 mL

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III. Sorvall WX 80+ Ultracentrifuge and Rotor Log Sheet

From: To:

Date Run Length Rotor Used RMP User Comments

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 IV. Acceleration and Deceleration Modes

The acceleration and deceleration rates can be adjusted between 0 to 500 rpm. By setting an
ideal acceleration and deceleration time samples will not be disturbed, which improves
efficiency and reduces centrifugation time. The table below shows the relationship between
ACCEL and DECEL code numbers and acceleration and deceleration time.

*1 Minimum time: The time it takes the drive motor to accelerate and decelerate the rotor using
maximum torque. This time will vary with rotor type and the mechanical
resistance of the drive motor.

*2 DECEL code "F": A deceleration method that does not involve brake control using the drive
motor. The deceleration time will then vary widely depending on
differences in the small mechanical resistance of the drive motor and the
vacuum level of the rotor chamber and other factors.

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V. Operation Flowchart

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 VI. Troubleshooting Table

Rotor Problems

Problem Probable Cause Solution

Balance the rotor’s load and

Rotor is unbalanced. distribute the weight
symmetrically.

The speed selected is near or

above the rotor’s critical Select a lower rotation speed.
speed range.
Severe Vibration
The rotor is incorrectly Verify the rotor’s assembly. Test
mounted. that it is well adjusted.

Lubricate the pivoting axis

There is a lack of lubrication
according to the manufacturer’s
in the rotor’s supports.
recommendation.

Open the ventilation line in the

A vacuum is being produced
upper part of the rotor or bucket
during centrifugation.
to eliminate the vacuum.
Rotor covers, canister or
cubes difficult to loosen Perform routine cleaning of the
after centrifugation. The rings are contaminated
rings and lubricate. Use products
with filth, dried lubricants or
recommended by the
metallic particles.
manufacturers.

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 Tube Problems

Problem Probable Cause Solution

The covers are badly secured. Adjust the covers.

The meniscus must be lower in

The tubes are too full.
order to prevent leaks.

The maximum recommended Verify the volume and speed

The tubes leak. level has been exceeded in the recommendations for the
open tubes. centrifugation.

Press lightly, after heat-sealing

A deficient seal is presumed (only if the contents are not
in the rapid seal tubes. affected). If leaks are visible, seal
again.

If the sample is frozen, warm to 2

The tubes can be broken or
°C before centrifuging. Evaluate
become fragile if they are used
how the tubes behave at low
below the recommended
temperatures before
temperature.
The tubes are cracked or centrifuging.
broken.

The tubes become fragile with Discard expired tubes, use new
age and use. ones.

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 Various Problems

Problem Probable Cause Solution

The main switch is in the

on position but the There is no power to the
Verify the power supply.
centrifuge is not instrument.
functioning.

The centrifuge cover Turn the centrifuge ON. Press

The centrifuge is off.
cannot be opened. the handle and open the cover.

The load to be centrifuged is

Balance the load to centrifuge.
The balance indicator is unbalanced.
activated.
The centrifuge is not levelled. Level the centrifuge.

The rotor adjustment Correctly adjust the fastening

mechanism is slack. system.

There is a vibration at Verify the balance of the load to

The load is unbalanced.
low speed. be centrifuged.
Select a more elevated rotation
The selected speed is close to
speed or use a different type of
the rotor’s resonance point.
rotor.

Turn off the centrifuge. Verify

There are fluctuations in The transmission belts are in
the condition and state of the
the rotation speed. a bad condition
belts.

Turn off the centrifuge. Verify

the condition of the brushes. If
The brushes are defective. this is the problem, put new
The rotation speed does
brushes with the same
not reach the selected
specifications as the originals.
speed.
The speed control calibration Adjust the speed control
is maladjusted. calibration.

The chamber is cold but The temperature is incorrectly

Verify the temperature selection.
the rotor is warm. selected.

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VII. Inspection, Maintenance and Service Record

Description of
Date Service Agent Signature
Action

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VIII. Documentation of Training (Signature of all users is required)

I, the undersigned, verify that I have read and understand the content and
responsibilities of this SOP and have undergone training by an approved
facility personnel.

Date Trainee Name Instructor Signature

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