Bridges Lab Protocols - User contributions [en]

Triglyceride Assay from Cells and Tissues

2019-10-11T17:03:31Z

Kkistler:

==Materials==
* '''Homogenization Buffer''' (50 mM Tris pH 8, 5 mM EDTA, 30 mM Mannitol, PI inhibitor, can be made in bulk without the PI, PI added fresh) (to make 50 mL of homogenization buffer - 2.5 mL of 1M Tris pH 8, 500 uL of 0.5 M EDTA, 0.273 g Mannitol, fill up the rest with water)
* 10M KOH (28.1g in 50 mL of water)
* '''Chloroform/Methanol Mixture''' (2:1)
* '''Butanol Mixture''': 3 mL butanol, 1.66 mL Triton-X114, 0.33 mL Methanol
* Sigma Triglyceride Assay Kit (Cat TR0100)

==Protocol==
#Weigh out 30-50mg tissue (record weights for normalization) on dry ice into round bottom eppendorf tube (2 mL). Add one stainless steel ball bearing.
#Add 500ul Homogenization Buffer
#Homogenize with Qiagen Tissue Lyser for 3 minutes @ 25Hz for Liver/WAT or for 5 minutes @ 30Hz for muscle
#Add 12.5ul KOH
#Mix by inverting then transfewr sample to a new 1.5 mL tube. Place dirty ball bearing in ethanol (located in the fume hood)
#Add 800ul '''Chloroform/Methanol Mixture'''
#Vortex vigorously then sit at room temperature for 5 minutes
#Centrifuge for 10 minutes @ 13000G
#Transfer 200 ul of the bottom layer into a new tube
#Centrifuge again for 7-10 minutes @ 13000G
#Transfer 150 ul of the bottom layer into the new tube (if there is enough of the bottom layer, transfer a total of 200 ul)
#Let evaporate overnight at room temperature
#Add'''(50ul)''' of '''Butanol Mixture''' and vortex. See Suggested Volumes for your specific tissue.
#Measure triglyceride levels using SIgma Diagnostic Kit, using 5ul of sample.
##Resuspend triglyceride and glycerol reagent with water if necessary
##Calculate how many sample you have (samples + blank + standard curve)
##Prepare reagent. You need 80uL of glycerol reagent and 20uL of triglyceride reagent. Make extra and combine in a Falcon tube.
##Aliquot '''100ul into a well of a 96 well plate'''
##For standards, add 0-5 and .5ul of glycerol standard
##Add 5ul of resuspended lipid to each well to start (also make a 5ul blank of the butanol mixture) and mix.
##Pop any bubbles with tip before incubating.
##Let sit for ~30 mins @ room temperature (or 5mins @ 37C if you are in a hurry). If using >10ul of butanol mix the solution may be cloudy. Let it settle and it should become more clear.
##Measure absorbance @ 540nm
##If any samples are A540<0.1 or above the 5ul standard the A540, then repeat with more or less lipid as required.

Triglyceride Assay from Cells and Tissues

2019-10-01T18:23:23Z

Kkistler:

==Materials==
* '''Homogenization Buffer''' (50 mM Tris pH 8, 5 mM EDTA, 30 mM Mannitol, PI inhibitor, can be made in bulk without the PI, PI added fresh) (to make 50 mL of homogenization buffer - 2.5 mL of 1M Tris pH 8, 500 uL of 0.5 M EDTA, 0.273 g Mannitol, fill up the rest with water)
* 10M KOH (28.1g in 50 mL of water)
* '''Chloroform/Methanol Mixture''' (2:1)
* '''Butanol Mixture''': 3 mL butanol, 1.66 mL Triton-X114, 0.33 mL Methanol
* Sigma Triglyceride Assay Kit (Cat TR0100)

==Protocol==
#Weigh out 30-50mg tissue (record weights for normalization) on dry ice into round bottom eppendorf tube (2 mL). Add one stainless steel ball bearing.
#Add 500ul Homogenization Buffer
#Homogenize with Qiagen Tissue Lyser for 3 minutes @ 25Hz for Liver/WAT or for 5 minutes @ 30Hz for muscle
#Add 12.5ul KOH
#Mix by inverting then transfewr sample to a new 1.5 mL tube. Place dirty ball bearing in ethanol (located in the fume hood)
#Add 800ul '''Chloroform/Methanol Mixture'''
#Vortex vigorously then sit at room temperature for 5 minutes
#Centrifuge for 10 minutes @ 13000G
#Transfer 400 ul of the bottom layer into a new tube
#Let evaporate overnight at room temperature
#Add'''(50ul)''' of '''Butanol Mixture''' and vortex. See Suggested Volumes for your specific tissue.
#Measure triglyceride levels using SIgma Diagnostic Kit, using 5ul of sample.
##Resuspend triglyceride and glycerol reagent with water if necessary
##Calculate how many sample you have (samples + blank + standard curve)
##Prepare reagent. You need 80uL of glycerol reagent and 20uL of triglyceride reagent. Make extra and combine in a Falcon tube.
##Aliquot '''100ul into a well of a 96 well plate'''
##For standards, add 0-5 and .5ul of glycerol standard
##Add 5ul of resuspended lipid to each well to start (also make a 5ul blank of the butanol mixture) and mix.
##Pop any bubbles with tip before incubating.
##Let sit for ~30 mins @ room temperature (or 5mins @ 37C if you are in a hurry). If using >10ul of butanol mix the solution may be cloudy. Let it settle and it should become more clear.
##Measure absorbance @ 540nm
##If any samples are A540<0.1 or above the 5ul standard the A540, then repeat with more or less lipid as required.

Spill Response Procedures for Infectious agents and Recombinant DNA

2019-09-13T19:50:49Z

Kkistler: KK signed

Spills and accidents should be immediately reported to the Principal Investigator and the Department of Occupational Safety and Environmental Health (OSEH) (647-1143).

Large Spills Outside of the Biosafety Cabinet
OSEH will respond to large spills (greater than 200 ml) of BSL-2 (or above) containment level material. Contact OSEH (647-1143) for help with large spill cleanup.
Small Spills Outside of the Biosafety Cabinet
# Don double gloves and eye protection if not already wearing them.
# Cover spilled material with an absorbent paper towel or Kimwipe. Once the absorbent material is in place over the spill, wet the material with a 10% solution of bleach (1:10 dilution of Clorox containing sodium hypochlorite) or other appropriate disinfectant.
# Let stand 15-20 minutes, wipe up and wash surface with appropriate disinfectant.
# Wipe down all equipment and surfaces which may have been splashed.
Spill in a Biosafety Cabinet
Note: Leave the cabinet turned on.
# Don double gloves, a lab coat, and eye protection if not already wearing them.
# Cover spilled material with an absorbent paper towel or Kimwipe. Once the absorbent material is in place, wet material with 10% solution of bleach (1:10 dilution of Clorox containing sodium hypochlorite) or other appropriate disinfectant. Let stand 15-20 minutes, wipe up and wash surface with appropriate disinfectant.
# If personnel are contaminated, remove potentially contaminated garments at the BSC and decontaminate garments by saturation with 70% ethanol or place in autoclave bag for autoclaving. Wash hands and other potentially exposed skin surfaces thoroughly with soap and water. Don fresh PPE, return to worksite, and spray walls, liners, and equipment with an appropriate disinfectant.
# Spray or wipe container walls, other work surfaces and equipment with the appropriate disinfectant
# If necessary, flood the work surface, drain pan and catch basin below the work surface with disinfectant. Allow at least 15-20 minutes contact time.
# Soak up the disinfectant and drain the catch basin into a container. Lift the front exhaust grille and tray and wipe all surfaces. Ensure that no foreign materials are blown into the area below the grille.
# If a 10% bleach solution is used on metal surfaces, rinse with water or 70% ethanol after decontamination is complete.
# If the spill overflows into the interior of the cabinet, more extensive decontamination of the cabinet may be necessary. Contact OSEH (763-6973) for decontamination of the cabinet.
Spill in the Laboratory
# If an infectious aerosol may exist outside of a biosafety cabinet, leave the room immediately, lock the door, post a warning sign and inform your supervisor. If clothing is contaminated, remove and turn the exposed side of fabric in on itself and place in autoclave bag or biohazard container. Wait at least 30 minutes before reentering the lab to allow dissipation of aerosol created by the spill. During this time, review clean-up procedures, assemble decontamination materials, PPE and contact OSEH (647-1143).
# Don fresh gloves, a lab coat or tyvek suit, and eye protection.
# Carefully lay disinfectant-soaked towels over the spill and pour more around the spill. Use more concentrated disinfectant if the volume of material will significantly dilute the disinfectant.
# Allow 15-20 minutes contact time.
# Use forceps to place sharp objects into a sharps container. Using a dustpan and dustbroom, tongs, etc., transfer all contaminated materials (paper towels, gloves, labware, etc.) biohazard waste containers and contact OSEH HazMat (763-4568) for removal.
# Wipe surrounding surfaces with disinfectant to cover all splash areas. Wipe flat surfaces to remove any material that may have splashed out and settled on those surfaces.
# Place all contaminated materials, including protective clothing, into an autoclave bag or biohazard waste container.
# Wash hands with soap and warm water.
# If a personnel exposure occurred, complete an Illness or Injury Report form (https://www.workconnections.umich.edu/illnessorinjury.php) summit to Work~Connections and report incident to OSEH (763-6973). See additional procedures noted below.
Skin, Mucous Membrane, or Injury Exposure
# If there is an agent-specific protocol for exposures, follow that (e.g., HIV, Herpes B).
# In the case of skin contact or injury with a contaminated instrument:
## Thoroughly wash area with soap and water. Do not squeeze the wound to induce bleeding.
## Avoid use of abrasive chemical soaps or disinfectant washes as they can cause skin abrasions and a possible additional route of entry for the agent.
## For mucous membranes (e.g., eyes, mouth), flush for a minimum of 15 minutes.
# Notify UM-Occupational Health Services (OHS) for evaluation of exposure following:
## Contact with mucous membranes;
## Contact with non-intact skin;
## Percutaneous exposure; or
## Any type of exposure that involves concentrated virus.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:44, 13 September 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:28, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:58, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:21, 12 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 17:26, 15 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 21:43, 12 September 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges
Revision Date - 2016-09-13

[[ Category:SOP ]]

SOP - Formalin and Paraformaldehyde

2019-09-13T19:47:57Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

==Description==
This standard operating procedure outlines the handling and use of formalin and paraformaldehyde. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling formalin and paraformaldehyde.

Formalin and paraformaldehyde are primarily used as fixatives. These fixatives act to preserve and stabilize cells and tissues prior to examination processes. The aqueous solution of formalin is 37-40 percent formaldehyde in water or methanol. Paraformaldehyde is the crystallized polymer of formaldehyde (97%) that is weighed out and dissolved in solution for experimentation or for cell and tissue fixation. Typically 3-10% formalin or paraformaldehyde solutions are used to perfuse or fix tissues.

==Useful Formalin and Paraformaldehyde Links:==
http://www.cdc.gov/niosh/docs/81-111/

http://www.cdc.gov/niosh/topics/formaldehyde/

http://www.cdc.gov/niosh/npg/npgd0293.html

==Potential Hazards==
*Formalin and paraformaldehyde solutions can emit formaldehyde gas, a known human carcinogen, and can irritate the eyes and skin.
*Working with paraformaldehyde powder (and, to a lesser extent, flakes or granules), can expose employees to paraformaldehyde dust, which is a strong irritant/sensitizer.
*Contact with these solutions or paraformaldehyde solids may also cause drying of the skin and/or allergic dermatitis.
*The MIOSHA Permissible Exposure Limit for formaldehyde is 0.75 ppm for 8 hours or 2 ppm for 15 minutes. There is a substance-specific MIOSHA standard for formaldehyde, and an action limit of 0.5 ppm.
*Consult your Safety Data Sheet for more information on hazards.

==Engineering Controls==
*Work with concentrated (>4% formaldehyde/paraformaldehyde) solutions only in a chemical fume hood.
*Handle paraformaldehyde powder (and, preferably, granules or flakes) only in a chemical fume hood.
*Dilute solutions (<4% formaldehyde) may be used on the benchtop in small quantities.
*If there is any possibility that an employee's eyes may be splashed with solutions containing 0.1 percent or greater formaldehyde, an eyewash/drench hose must be available within the immediate work area for emergency use.
*If employees' skin may become splashed with solutions containing 1 percent or greater formaldehyde, for example, because of equipment failure or improper work practices, the MIOSHA formaldehyde standard requires a conveniently-located safety shower. Contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 to determine if a safety shower will be needed.

==Work Practice Controls==
*Designate an area for working with concentrated formalin, concentrated paraformaldehyde solutions, and paraformaldehyde solid, and label it as such.
*Keep containers closed as much as possible.
*Use in the smallest practical quantities for the experiment being performed.
*If you are weighing paraformaldehyde powder and the balance cannot be located in a fume hood or BSC, tare a container then add powder in the hood and cover before returning to the balance to weigh the powder.
*Labs handling moderate to large quantities of formaldehyde-containing solutions on a regular basis should contact OSEH at (734) 647-1143 for assessment of exposure. Areas that handle only small (100 ml or less) pre-filled specimen containers, or that work with formaldehyde-containing solutions exclusively in a functioning chemical fume hood, would have low potential for overexposure, but should contact OSEH if there are concerns.
*Once work with formalin/paraformaldehyde is complete, wipe down area with a soap and water solution.

==Protective Equipment==
Wear standard nitrile laboratory gloves, chemical splash goggles, face shield, and lab coat. If splash may occur, also wear an impervious apron. (MIOSHA requires that all contact of the eyes and skin with liquids containing 1 percent or more formaldehyde be prevented by the use of chemical protective clothing made of material impervious to formaldehyde and the use of other personal protective equipment, such as goggles and face shields, as appropriate to the operation.)

==Transportation and Storage==
*Transport formaldehyde solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
*Keep container in cool, well-ventilated area.
*Keep container tightly closed and sealed until ready for use.
*Store in secondary containment with flammables, away from oxidizers, reducing agents, metals, and acids.
*Keep containers of paraformaldehyde (PFA) solid away from water.
*Avoid storing on the floor.
*Avoid ignition sources.

==Waste Disposal==
Formalin and paraformaldehyde solutions and powders must be disposed following the guidelines above while accumulating wastes and awaiting chemical waste pickup. Waste must be disposed of following OSEH Hazardous Materials procedures. Contact OSEH’s Hazardous Materials Management (HMM) at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.
'''
For an actual chemical exposure/injury,
*Flush exposed eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
*If there is respiratory irritation associated with exposure, remove all persons from the contaminated area.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedure==
Employees in the area should be prepared to clean up minor spills, including most spills confined to the chemical fume hood. Wearing double nitrile gloves, splash goggles, face shield and lab coat (and impermeable apron, if available), use absorbent pads to absorb spilled material. For small spills of solid PFA, dampen the absorbent pad with methanol before placing over the spilled material and allow to sit for a few minutes before wiping up. After spill has been completely absorbed, wash down contaminated area with soap and water at least two times. Contaminated PPE and clean-up materials must be placed in a clear plastic bag or compatible container for pick-up by OSEH.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

For large spills and most spills of formalin or paraformaldehyde solutions or paraformaldehyde powder that occurs outside of a chemical fume hood should be referred to OSEH-HMM.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling formalin and paraformaldehyde.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 15:58, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:23, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:31, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:50, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:49, 13 June 2017 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Sensitizers

2019-09-13T19:44:40Z

Kkistler: KK signed

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of sensitizers. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling sensitizers.

==Potential Hazards==
A sensitizer (allergen) is a substance that causes exposed people to develop an allergic reaction in normal tissue after repeated contact with the substance. The reaction might be as mild as a rash (contact dermatitis) or as serious as anaphylactic shock. Sensitizers may also be corrosive or carcinogenic.

Examples of compounds that may cause sensitization in some individuals are diazomethane, various isocyanates, formaldehyde, latex, and benzylic and allylic halides.

==Engineering Controls==
Use a properly functioning lab fume hood when handling sensitizers that can be inhaled (via mist/fume/gas/vapor).
If the process does not permit the handing of such materials in a fume hood, contact OSEH at (734) 647-1143 for review of the adequacy of ventilation measures.

==Work Practice Controls==
For sensitizers that are also considered a particularly hazardous substances, a designated area shall be established per the other applicable SOP.
*The use of latex gloves may sensitize you to latex. If you feel that you develop a reaction to latex, please inform the PI and nitrile gloves will be provided for you.
*The use of masks while in the animal rooms may help prevent sensitizing to animal dander. These are provided in each animal room and are highly recommended for use each time interacting with the animals.

==Personal Protective Equipment (PPE)==
At minimum, safety glasses, lab coat, long pants, and closed toed shoes are to be worn when entering laboratories having hazardous chemicals.

Additionally, when handling hazardous chemicals or contacting potentially contaminated surfaces, protective gloves are to be worn. For proper selection of glove material, review the SDS or refer to the glove compatibility charts available through the OSEH website.

For hazardous chemicals that are toxic via skin contact/ absorption, additional protective clothing (i.e., face shield, apron, oversleeves) is appropriate where chemical contact with body/skin is foreseeable.

Goggles (not safety glasses) are appropriate for processes where splash or spray is foreseeable.

==Transportation and Storage==
Follow any substance-specific storage guidance provided in the SDS documentation.
�
==Waste Disposal==
Many sensitizers intended for disposal may likely be considered hazardous wastes. For general guidance regarding waste disposal, refer to the OSEH Hazardous Materials Management (HMM) chemical waste information available online. For specific guidance, contact OSEH-HMM at (734) 763-4568.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact Occupational Health Services (734) 764-8021 or the UMHS Emergency Room (734) 936-6666 for advice. Be prepared to provide them with a copy of the SDS so that medical personnel are aware of the material(s) involved and take precautions to protect themselves.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==TREATMENT FACILITIES:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

==Spill Procedure==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

For a small spill, inside the fume hood, use proper personal protective equipment as indicated above. Absorb the material with an absorbent, non-combustible material such as sorbent pads, sand or vermiculite. Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

==MAJOR CHEMICAL SPILL==
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling sensitizers.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:13, 19 October 2016 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:55, 7 November 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:27, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:53, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:22, 12 June 2017 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Phenol

2019-09-13T19:42:11Z

Kkistler: KK signed

[[Category: SOP]]
[[Category: Lab Safety]]
'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure'''

=Description=
This standard operating procedure (SOP) outlines the handling and use of phenol. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling phenol.

Phenol is used as a reagent in chemical analysis. It causes burns and risk of serious damage to eyes. Phenol is toxic by inhalation, in contact with skin and if swallowed.

=Potential Hazards=
• Phenol is very hazardous in case of skin contact – it is corrosive, toxic, and can cause irritation. Initially, it can cause numbness or slight tingling, so employees may not be immediately aware of contact. If absorbed through the skin it can cause muscle weakness, tremors, loss of coordination, shock, sudden collapse, coma, convulsions, organ damage, and death. It may cause severe eye injury (including blindness) if it contacts the eyes, and is extremely toxic (fatal) by ingestion.

• Inhalation exposure is less likely – it does not evaporate easily at room temperature, but can be inhaled if heated and/or misted, or in the case of a large spill. If inhaled, phenol can cause upper respiratory irritation, lung damage, and CNS impairment.

• For more safety information, refer to Prudent Practice’s Laboratory Chemical Safety Summary for Phenol.

Occupational Exposure Limits (OELs):
• MIOSHA PEL and ACGIH TLV: 5 ppm, 8-hour TWA

=Engineering Controls=
An eyewash (preferably eyewash/drench hose combination unit) must be located in the immediate area. If medium-large quantities are used (>~100 ml of 100% phenol), a safety shower is also required. Contact OSEH at (734) 647-1143 for an assessment of the need for a safety shower if one is not available. Work with large open containers should be performed only in a chemical fume hood. Small amounts can be handled safely on the bench top, as long as skin contact is avoided.
�
=Work Practice Controls=
• Designate areas where phenol is stored or manipulated.

• It is highly recommended that labs using phenol (or reagents containing phenol – e.g., TRIzol) have polyethylene glycol 300 or 400 (PEG-300 or PEG-400) on hand in case of dermal exposure.
• Purchase in the smallest container that is practical for lab use.

• Purchase in a shatter-resistant container if available (such as PVC-coated glass).

• Keep containers closed as much as possible.

• Use in the smallest quantities and lowest concentration practicable for the experiment being performed.

• After work with phenol is complete, wipe down work area with soap and water solution.

=Personal Protective Equipment=
• Wear neoprene or doubled nitrile laboratory gloves, safety glasses, and lab coat. If gloves are splashed, remove them immediately and put on a new pair of gloves. If splashes may occur, wear a face shield with chemical splash goggles, and an impervious apron over the lab coat.

• If phenol and chloroform will be used together, please note that chloroform easily penetrates nitrile gloves and can degrade them, allowing phenol to contact the skin. Plan work to avoid glove contact and change gloves immediately if there is a splash. If extensive work with phenol and chloroform is done in the lab, consider the reusable ChemTek Viton/Butyl glove made for work with both of these chemicals (or they can be used for spill clean-up).

=Transportation and Storage=
• Transport phenol in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Keep container in cool, well-ventilated area.

• Keep container tightly closed and sealed until ready for use.

• Store in secondary containment away from moisture, strong oxidizers, strong caustics, plastics, rubber, nitric acid, water + heat, and chemically active metals, such as aluminum and magnesium powder, sodium, potassium, and lithium.

• Store below eye level.

• Avoid storing on the floor.

• Avoid ignition sources.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.
�
=Exposures/Unintended contact=

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
• Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).

• If a small area of skin has been exposed to phenol, swab polyethylene glycol 300 or 400 (PEG-300 or 400) onto the affected area immediately and repeatedly until the smell of phenol is no longer evident, then seek medical attention. If PEG-300 or 400 is not available, flush area with COPIOUS amounts of water (such as from a drench hose or safety shower) for at least 15 minutes, then seek medical attention. See the US Agency for Toxic Substances and Disease Registry (ATSDR), Medical Management Guidelines for Phenol.

• For larger areas of exposed skin or eye exposure, flush area with COPIOUS amounts of water for at least 15 minutes, and then seek immediate medical attention. Please note that using high-density water irrigation will reduce phenol uptake, but if lesser amounts of water are used it will merely dilute the phenol and increase the area of exposure.

• For situations with risk of inhalation exposure (including spills of powder outside of a chemical fume hood), remove all persons from the contaminated area.

• If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

�
=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

• Employees in the area should be prepared to clean up minor spills, including most spills confined to the chemical fume hood. Wearing double nitrile gloves, splash goggles, face shield and lab coat (and impermeable apron, if available), use absorbent pads to absorb spilled material. After spill has been completely absorbed, wipe down contaminated area with soap and water solution. Lab personnel should take great care to avoid skin contact with phenol. If skin contact does occur, follow the instructions outlined for exposures/unintended contact in the section above.

• Contaminated PPE and clean-up materials should be placed in a clear plastic bag. Call OSEH-HMM (734) 763-4568 to arrange for waste pick-up.

• If a large spill occurs, leave the lab and call University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance from OSEH. Do not take any action to cover the spill. Post a warning on the lab and do not allow others to enter. Have a person available that has knowledge of the incident and laboratory to assist emergency personnel.

Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

Training of Personnel
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling phenol.

Certification
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:50, 10 November 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 21:08, 6 December 2016 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:47, 13 June 2017 (UTC)
*[[User:Cdgleason|Cdgleason]] ([[User talk:Cdgleason|talk]]) 03:55, 4 October 2018 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 21:43, 12 September 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - Needle Capping

2019-09-13T19:34:32Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This procedure outlines the methods for the safe recapping of needles and covers the safe handling, practices, disposal, and needlestick reporting requirements pertaining to hypodermic needles, hypodermic needles with syringes, needles with attached tubing, and other needle equipment. It is the responsibility of all laboratory staff to contribute to and comply with these procedures.

==Potential Hazards==
Needles present serious health hazards because people can be easily exposed parenterally to hazardous chemicals and infectious biological agents through injection. Recapping needles is extremely dangerous because it can result in the accidental stabbing of hands and other body parts. If working with human blood or other potentially infectious material, ensure that an Exposure Control Plan is completed and available. All applicable work requirements such as SOPs, universal precautions, hepatitis vaccination, etc. are required to be followed.

==Engineering Controls/Workplace Controls==
NEEDLES SHOULD NOT BE RECAPPED, BENT, REMOVED OR OTHERWISE MANIPULATED BY HAND.

* However, if it is essential that a needle be recapped due to the nature of the work, the use of a mechanical device or the one-handed scoop method must be used. In addition, there needs to be written justification and instructions for any procedures that involves the recapping of needles (SOP).

==Needle Recapping Instructions:==
1. One-Handed Scoop Method

Step 1: Place the cap on the desk or other flat surface with something firm to “push” the needle cap against.

Step 2: Holding the syringe with needle attached in one hand, slip the needle into the cap without using the other hand.

Step 3: Push the capped needle against a firm object to “seat” the cap onto the needle firmly using only one hand.

2. Needle Recapping Devices

These are devices with a hole in the center that hold needle caps and allow for single hand uncapping and recapping.

==Additional Guidelines==
* Keep needles away from fingers and other body parts.
* Place uncapped needles in a rigid tray during procedures and transportation.
* Do not leave unprotected needles on work surfaces.
* Shearing or breaking of contaminated needles is prohibited.
* Use disposable needle locking syringe units and other engineered sharps injury protections, which has a built-in safety feature or mechanism that effectively reduce exposures, and needleless systems whenever possible.
* Blunt cannulas should be used as alternatives to needles wherever possible.
* Needles and syringes should never be used as a substitute for pipettes.
* Used disposable needles and syringes must be placed in conveniently located appropriate sharps disposal containers and discarded as infectious waste.
* Use extreme caution and be alert when handling sharps. Avoid distractions and keep sight of the needle.
* Ensure that animals are restrained when using needles on animals and be on the alert for any unexpected movements.
* The use of needles and syringes should be restricted to procedures for which there is no alternative.
* Refer to the OSEH Exposure Control Plan and Biosafety Manual for additional information.

==Personal Protective Equipment (PPE)==
Wear safety glasses/goggles, gloves, lab coat/gown, close-toe shoes, and any other applicable PPE when handling needles and syringes.

==Waste Disposal==
Upon completion of a procedure involving use of needle equipment, immediately discard the items, including syringes connected to the needle, tubing, etc., into a clearly and properly labeled or color-coded rigid, closable, leakproof, puncture-resistant, disposable sharps container located in the work area at the point of use. All used needle equipment waste must be discarded as infectious waste and in compliance with OSEH requirements. NEVER reach into the sharps container. Never empty the contents into another container. Do not overfill the sharps container.

==Accidental Needle Stick/Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

If a needlestick injury should occur, wash the puncture area with soap and water, report the incident to the supervisor and principal investigator, and seek medical attention as soon as possible. Any needlestick with recombinant DNA and/or an infectious agent must be reported to the U-M Biological Safety Officer (BSO) (763-3133) as soon as possible. The U-M BSO will determine if institutional notification to Federal Agencies is required within 24 hours per applicable regulations. Failure to report may result in severe penalties to the researcher, laboratory, department or institution.
Report all needlesticks to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Please include (1) the type and brand of device involved in the incident. (2) The work unit or work area where the exposure incident occurred. (3) An explanation of how the incident occurred. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

In addition, as required for all lab incidents, complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for additional accident and injury reporting information.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling needles.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab Manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:24, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:33, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:03, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:00, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:50, 13 June 2017 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-14

SOP - Cryogenic Materials

2019-09-13T19:31:53Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure (SOP) outlines the handling and use of cryogenic materials. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling cryogenic materials.

Cryogenic liquids have various uses in the laboratory including sample preservation, cooling of equipment, and handling and storage of biological samples.

=Potential Hazards=
Cryogenic materials are liquefied gases with boiling points less than -73°C (-100°F). They can cause tissue damage (frostbite) and asphyxiation due to oxygen displacement. They also pose a fire hazard if oxygen is condensed out of the atmosphere during the use of cryogenic liquids that have boiling points that are lower than oxygen – e.g. nitrogen, helium or hydrogen. There is a potential for explosion due to pressure buildup in closed containers. Materials intended for use with cryogenic liquids must be selected carefully as they can be weakened or fail due to embrittlement.

=Engineering Controls=
Use and store cryogens in well-ventilated areas. Closets, small rooms, and walk-in cold rooms (or similar small/unvented spaces) should be avoided to prevent buildup of the gas as the cryogen evaporates/sublimes. If using large quantities of cryogens or if there are concerns about the adequacy of ventilation, consult OSEH Research Health and Safety (734) 647-1143 to determine if additional ventilation or an oxygen sensor/alarm is needed.

Each part of a cryogenic system must have its own pressure relief system.

=Work Practice Controls=
• Cryogenic gases can condense oxygen out of the air, creating a localized oxygen enriched environment. Therefore they must be used and stored away from flammable materials, open flames and ignition sources.

• Do not put your head inside a liquid nitrogen freezer or other enclosed space containing a cryogen. Never allow people to travel in an elevator with cryogenic liquids.

• Do not use hollow rods or tubes as dipsticks. (When a warm tube is inserted into a cryogen, liquid will spout from the top of the tube.

• When retrieving cryopreservation vials that have been stored in liquid nitrogen, raise the vials out of the liquid phase and store them in the gas above the liquid for 24 hours to allow any trapped liquid nitrogen to escape. After this period, the vials may be removed. Retrieving these vials directly from the liquid phase may cause the vials to rupture if liquid nitrogen has become trapped inside. This may expose the employee to flying bits of plastic, biological specimens or cryoprotective agents (such as dimethyl sulfoxide) that were in the vial.

• Check the liquid levels of Dewars regularly. If the liquid evaporates more rapidly than normal, the Dewar may be losing its vacuum.

• Also follow the SOP for asphyxiants.

=Personal Protective Equipment (PPE)=
• Wear loose fitting cryogenic gloves, safety goggles, face shield, lab coat, long pants and closed toe shoes when handling and transporting cryogenic materials.

• Lab coats should be knee length and have no pockets or cuffs to catch the liquid.

• Pants should not have cuffs and should be long enough to go over the shoe or boot.

=Transportation and Storage=
• Never store cryogenic liquids or dry ice in walk-in cold rooms.

• Use and store liquid hydrogen and helium away from flammable materials and ignition sources. (These gases can condense oxygen out of the air, creating a localized oxygen enriched environment.)

• For liquid helium and hydrogen storage systems, check the pressure relief and inspect the system for leaks regularly.

• These gases are cold enough to solidify atmospheric air; leaks in storage systems for these gases may become plugged with solidified air. If the pressure relief device becomes plugged, the container may over-pressurize and fail.

• Use and store liquid oxygen away from open flames. Post a “No Open Flames” sign in liquid oxygen storage areas.

• Store and transport cryogenic materials only in Dewars or cryogenic liquid cylinders designed for that particular cryogen. Inspect Dewars daily to insure that no air or ice plugs exist in the neck openings.

• Cryogens may be transported in elevators only in containers certified to leak at less than or equal to 1 liter of liquid (or 1 kilogram of solid) per day. Never allow passengers to travel in an elevator with cryogenic liquids.

=Waste Disposal=
If a cryogen (such as dry ice) has been mixed with a liquid that would normally be disposed of as hazardous waste (such as alcohol, as in a cold trap), follow disposal procedures for that liquid. • Cryogens must not be dumped into sinks, as the thermal shock is likely to damage the sink and/or the plumbing.

• As the unneeded cryogen is evaporating, good ventilation is essential to prevent hazardous air concentrations of the gas.

• Do not put unneeded cryogen in a sealed container, as the build-up of pressure could cause an explosion.

=Exposures/Unintended Contact=

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
• Refer to SDS.

• Flush exposed eyes or skin with water with warm water for at least 15 minutes, then seek medical attention (see below).

• Do not rub affected area.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

=Spill Procedure=
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

Spills of cryogenic liquid can cause depletion of oxygen in the area. For spills over 1 liter (or smaller amounts in very small rooms), leave the area immediately and call the University of Michigan Division of Public Safety and Security (DPSS) at 911.
Additional Spill Links:
• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling cryogenic materials.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 20:44, 10 November 2016 (UTC)
*[[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:11, 12 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]])
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 21:43, 12 September 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☐ NO

Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - Chloroform

2019-09-13T19:28:33Z

Kkistler: KK signed

[[ Category: SOP ]]

==Description==
This standard operating procedure outlines the handling and use of chloroform. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling chloroform.

Chloroform (CAS # = 67-66-3) is a clear, colorless liquid with a pleasant, nonirritating odor that can be detected as low as 11.7 ppm (odor threshold) but more likely detected at levels of 85 to 307 ppm (well above safe exposure limits). It will burn only when it reaches very high temperatures.

NOTE: Stabilizers are often added to prevent oxidation by air and light, and to prevent chloroform from becoming acidic and corrosive.

Synonyms include: Trichloromethane, Methane trichloride, Formyl trichloride, Trichloroform, Methenyl trichloride and Freon 20.

Chloroform is used as an extractant solvent in manufacture of rubber, essential oils, sterols and alkaloids, guttapercha, resins, and in the recovery of fat from waste products; in chemical analysis and assays; and in photographic processing.
It is also used as a general solvent of lacquers, plastics, dyes, fats, greases, gums, oils, adhesives, and waxes, and in the rubber cleaning and dry cleaning industries.

==Useful Chloroform Links:==
* http://www.atsdr.cdc.gov/substances/toxsubstance.asp?toxid=16
* http://www.atsdr.cdc.gov/toxfaqs/tf.asp?id=52&tid=16
* http://www.nap.edu/openbook.php?record_id=4911&page=282

==Potential Hazards==
* Chloroform has been classified as a potential to probable human carcinogen, based on adequate evidence for carcinogenicity in animals. Some animal studies also show evidence of reproductive and developmental toxicity from chloroform exposure.
* Inhalation of vapors can cause headaches, drowsiness, dizziness, and nausea. At high concentrations disorientation, anesthetic effects, and unconsciousness can occur, but acute toxicity is low.
* Chloroform is an eye, skin and upper respiratory tract irritant.
* Chloroform is not combustible but exposure to fire or high temperatures may lead to formation of phosgene, hydrogen chloride and chlorine, all highly toxic gases.
* Consult the SDS for Chloroform for additional guidance and information.

Occupational Exposure Limits (OELs):
* MIOSHA: 2 ppm, 8-hour PEL

==Engineering Controls==
Work with chloroform in a chemical fume hood. Dilute solutions (< 10%) may be used on the benchtop in small quantities (< 500 mL).

==Work Practice Controls==
* Keep containers of chloroform closed as much as possible.
* Be aware of skin absorption as a possible route of exposure. Plan work so that minimal glove contact is expected, and purchase appropriate gloves for cleaning up small spills. (Refer to the PPE section below, for glove recommendations.)
* Use in the smallest practical quantities for the experiment being performed.
* Do not mix or store with acids; may form toxic gas.
* Thoroughly wash hands when finished handling.

==Personal Protective Equipment (PPE)==
* The minimum PPE for work with chloroform is Viton or PVA (Polyvinyl Acetate) laboratory gloves, lab coat, and safety glasses (ANSI/ISEA Z87.1 approved). Keep in mind that chloroform will penetrate nitrile gloves in less than 2.5 minutes.
* If a splash may occur, wear chemical splash goggles and/or a face shield.
* OSEH’s Glove Compatibility webpage can also be used to determine the recommended gloves.

==Transportation and Storage==
* Do not store chloroform with incompatibles. Chloroform is not compatible with the following: acetone, strong bases, alkalis, chemically-active metals (such as aluminum, magnesium, sodium, or potassium), dinitrogen tetroxide, fluorine, disilane, sodium methylate, triisopropylphosphine, and solid potassium tert-butoxide.
* Transport chloroform in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in secondary containment.
* Avoid storing on the floor.
* Refer to OSEH’s Chemical Storage webpage for additional information on the storage of hazardous materials.

==Waste Disposal==
Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. If breathing is difficult, give oxygen. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.
* Use proper personal protective equipment (PPE) as noted above.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., (less than 1 Gallon or 3.5 Liters). A major/large chemical spill requires active assistance from emergency personnel.

===Spill Response Steps:===

===MINOR CHEMICAL SPILL===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Use proper personal protective equipment as indicated above.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===MAJOR CHEMICAL SPILL===
Report large chemical spills greater than 1 Gallon or 3.5 Liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to U-M Division of Public Safety and Security (DPSS) by calling 911.
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Call U-M Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling chloroform.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:42, 15 August 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:15, 31 October 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:50, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:01, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:06, 12 June 2017 (UTC)
*[[User:Cdgleason|Cdgleason]] ([[User talk:Cdgleason|talk]]) 03:54, 4 October 2018 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-08-15

SOP - Centrifuge Safety

2019-09-13T19:23:54Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the use of centrifuges. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when using centrifuges.

Centrifuges are machines used to separate solids from liquids in a suspension. The spinning motion of a centrifuge produces centrifugal forces that separate substances of greater and lesser densities.

==Potential Hazards==
The centrifuge must be loaded carefully balanced. An unbalanced load may present a risk to both the machine and to persons nearby. Centrifuge rotors should never be touched while the rotor is spinning, as they present a significant personal risk when in motion. Another potential hazard is the possible aerosolization of harmful samples during centrifuge operation.

==Engineering Controls==
If centrifuging hazardous materials, care should be taken to use tightly capped tubes and/or sealable safety cups or rotors that can be loaded and unloaded in a fume hood or biosafety cabinet, depending on the hazard.

==Work Practice Controls==
The following information may be integrated into a lab-specific standard operating procedure (SOP) for centrifuge use.

1. ''Preventive Maintenance''

A. Establish preventive maintenance schedule: Including regular cleaning of centrifuge interior to prevent damage and avoid costly repairs. Reference centrifuge operator’s manual or contact manufacturer for guidance.

Equipment repair and adjustments shall only be conducted by qualified service technicians.

B. Maintain log book: For all high speed centrifuges and ultracentrifuges include run dates, durations, speeds, total rotor revolutions, and notes on rotor condition.

Retire rotors after manufacturer’s recommended life span except where annual stress test demonstrates absence of structural flaws. Note: Rotor life span may be reduced or warranty voided if autoclaved; contact manufacturer for guidance.

2. ''Planning for Use''

A. Complete lab-specific training for the centrifuge.

B. Wear appropriate PPE: Including safety eyewear, gloves, lab coat, and appropriate street clothing (i.e., long pants and closed-toe shoes). Ensure gloves are compatible with hazard(s).

C. Inspect centrifuge:
* Ensure tubes are rated for intended use (speed, temperature, and chemical resistance).
* Ensure rotor is compatible with centrifuge and seated on the drive correctly.
* Ensure rotor and safety cups/buckets are free of cracks and deformities.
* Ensure rotor O-ring is not cracked, missing, or worn.
* Ensure safety cups/buckets are attached correctly and able to move freely.

Contact a qualified service technician if inspection identifies centrifuge components requiring repair or replacement.

D. Prepare centrifuge tubes for loading:
* Inspect centrifuge tubes before use.
* Follow manufacturer’s filling limits for tubes. Do not overfill or underfill tubes.
* For biohazardous materials, disinfect outside of tubes prior to removal from biosafety cabinet and loading into rotor.

When centrifuging hazardous materials, use tightly capped tubes and/or sealable safety cups or rotors that can be loaded and unloaded in a fume hood or biosafety cabinet, depending on hazard. If a specific procedure does not allow for this, contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 for guidance.

E. Use in-line filter: For high speed centrifuges and ultracentrifuges, use in-line filters to prevent contamination of vacuum pump and pump oil. Provide secondary containment for vacuum pump.

3. ''Centrifuge Operation''

A. Balance centrifuge: Follow manufacturer’s instructions for proper centrifuge balancing steps.

B. Start run
* Do not leave centrifuge until full operating speed is reached and it appears to be running safely without incident.
* Stop centrifuge immediately if you notice any unusual noises or shaking. Confirm rotor is balanced.

To prevent rotor failure, do not exceed maximum speed and maximum mass limits for the rotor. You must reduce rotor speed if sample density calculations indicate maximum mass limits will be exceeded; contact manufacturer for guidance.

4 ''Sample Removal''

A. Stop run: Ensure centrifuge comes to complete stop before opening cover. When centrifuging hazardous materials, wait at least 10 minutes after run to allow aerosols to settle before opening centrifuge.

B. Check for leaks/spills: In samples, rotor, safety cups/buckets, and centrifuge well.

C. Open sealable tubes/safety cups/rotors: Wear appropriate PPE and open inside fume hood or biosafety cabinet, depending on hazard.

==Personal Protective Equipment==
When operating the centrifuge, proper eye protection (glasses/goggles), gloves, lab coat, and close-toe footwear should be worn. Gloves should be appropriate for the present hazard.

==Waste Disposal==
Centrifuge Disposal

1. For biohazardous materials, clean/disinfect centrifuge and remove/cross out biohazard sticker. Attach note on centrifuge describing what has been done.

2. For radioactive materials, request radiation safety survey and signage before disposal of centrifuge.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

TREATMENT FACILITIES:
U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
1. Mechanical Failure

Turn off centrifuge immediately and unplug power cord. Do not use centrifuge again until inspected by qualified service technician.

2. Hazardous Material Spill/Exposure

A. Turn off centrifuge immediately. Keep centrifuge cover closed for at least 30 minutes to reduce aerosolization of hazardous material.

B. Follow spill, exposure, and incident reporting instructions.
* For chemical, radioactive, and biohazard spills, contact OSEH at (734) 647-1143 for assistance and guidance.

Remember:
* Conduct rotor or safety cup/bucket cleanup in nearest biosafety cabinet or fume hood, depending on hazard.
* Use tongs or forceps to avoid contact with sharp-edged debris.
* Avoid alkaline cleaners for aluminum centrifuge components.
* Avoid abrasive wire brushes for cleaning.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling centrifuges.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]]) 17:19, 14 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:18, 10 November 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:49, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:01, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:59, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:50, 13 June 2017 (UTC)
*[[User:Nlambrec|Nlambrec]] ([[User talk:Nlambrec|talk]]) 21:16, 17 February 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-14

SOP - Carcinogens

2019-09-13T19:22:18Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

'''Laboratory Director (LD) Approval is Required Prior to Performing this Procedure
'''
==Description==
This standard operating procedure is designed to provide guidance in writing procedures for the safe handling and disposal of carcinogens, which are regarded as particularly hazardous substances. Review this document and supply the information required in order to make it specific to your laboratory.

A carcinogen is defined by meeting one the following:
*It is listed under the category, “known to be carcinogens” in the latest National Toxicology Program annual report.
*It is listed under Group 1 by the International Agency for Research on Cancer (IARC).
*It is listed in Group 2A or 2B by IARC or under the category, “reasonably anticipated to be carcinogens” by NTP, and causes statistically significant tumor incidence in experimental animals in accordance with any of the following criteria:
**After inhalation exposure of 6-7 hours per day, 5 days per week, for a significant portion of a lifetime to dosages of less than 10 mg/m3.
**After repeated skin application of less than 300 mg/kg of body weight per week.
**After oral dosages of less than 50 mg/kg of body weight per day.
If you have questions concerning the applicability of any item listed in this procedure contact the LD/Laboratory Supervisor or Occupational Safety and Environmental Health at (734) 647-1143.

==Potential Hazards==
A carcinogen commonly describes any agent that can initiate or speed the development of malignant or potentially malignant tumors, malignant neoplastic proliferation of cells, or cells that possess such material.

==Engineering Controls==
*Manipulation of carcinogens should be carried out in a fume hood. If the use of a fume hood proves impractical refer to the section on special ventilation.
*Certain carcinogens must be handled in a glove box rather than a fume hood. Occupational Safety and Environmental Health (734) 647-1143 or the LD will determine if this is required.
*Where the eyes or body of any person may be exposed to carcinogens, suitable facilities for quick drenching or flushing of the eyes and body shall be provided within the work area for immediate emergency use. Bottle type eyewash stations are not acceptable.
*Safety shielding is required any time there is a risk of explosion, splash hazard or a highly exothermic reaction. All manipulations of carcinogens which pose this risk should occur in a fume hood with the sash in the lowest feasible position. Portable shields, which provide protection to all laboratory occupants, are acceptable.
*Manipulation of carcinogens outside of a fume hood may require special ventilation controls in order to minimize exposure to the material. Fume hoods provide the best protection against exposure to carcinogens in the laboratory and are the preferred ventilation control device. When possible, handle carcinogens in a fume hood. If the use of a fume hood proves impractical, attempt to work in a glove box or on an isolated area on the bench top.
*If available, consider using a Biological Safety Cabinet. The biological safety cabinet is designed to remove particulates (the carcinogen) before the air is discharged into the environment. Carcinogens that are volatile must not be used in a biological safety cabinet unless the cabinet is vented to the outdoors.
*If your research does not permit the handling of carcinogens in a fume hood, biological safety cabinet, or glove box, you must contact Occupational Safety and Environmental Health (OSEH).
*Evacuated glassware can implode and eject flying glass, and splattered chemicals. Vacuum work involving carcinogens must be conducted in a fume hood, glove box or isolated in an acceptable manner.
*Mechanical vacuum pumps must be protected using cold traps and, where appropriate, filtered to prevent particulate release. The exhaust for the pumps must be vented into an exhaust hood.

==Work Practice Controls==
All areas where carcinogens are stored or manipulated must be labeled as a designated area.
*The room sign for the laboratory must contain a “Designated Area Within” identifier.
*All locations within the laboratory where carcinogens are handled should be demarcated with designated area caution tape (available from OSEH, or your department) and/or posted with designated area caution signs. This includes all fume hoods and bench tops where the carcinogens are handled.
*All employees with access to designated areas must be made aware of the substances being used and necessary precautions by the LD.
*Where feasible, carcinogens should be manipulated over plastic-backed disposable paper work surfaces. These disposable work surfaces minimize work area contamination and simplify clean up.

'''Doorways:''' The room sign must contain a “Designated Area Within, Caution” where carcinogens, reproductive hazards, and/or acutely toxic chemicals are stored or used.

'''Containers:''' All containers of carcinogens must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

Wash hands, forearms, face, and neck upon exit of a designated area to decontaminate.

Keep accurate records of amounts of carcinogenic substances being stored and used, including dates of use and names of researchers using the substances.

==Personal Protective Equipment==
Eye protection in the form of safety glasses must be worn at all times when handling carcinogens. Ordinary (street) prescription glasses do not provide adequate protection. (Contrary to popular opinion these glasses cannot pass the rigorous test for industrial safety glasses.) Adequate safety glasses must meet the requirements of the Practice for Occupational and Educational Eye and Face Protection (ANSI Z.87. 1) and must be equipped with side shields. Safety glasses with side shields do not provide adequate protection from splashes; therefore, when the potential for splash hazard exists, other eye protection and/or face protection must be worn.
Gloves must be worn when handling carcinogens. Disposable nitrile gloves provide adequate protection against accidental hand contact with small quantities of most laboratory chemicals. Lab workers should contact OSEH for advice on chemical resistant glove selection when direct or prolonged contact with hazardous chemicals is anticipated.
Lab coats, closed toed shoes and long sleeved clothing must be worn when handling carcinogens. Additional protective clothing should be worn if the possibility of skin contact is likely.

==Waste Disposal==
Because most spent, unused and expired chemicals/materials are considered hazardous wastes, they must be properly disposed of. Do not dispose of chemical wastes by dumping them down a sink, flushing in a toilet or discarding in regular trash containers, unless authorized by OSEH Hazardous Materials Management (HMM). Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.
'''

For an actual chemical exposure/injury:
*Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).
*For situations with risk of inhalation exposure (including spills of powder outside of a chemical fume hood), remove all persons from the contaminated area.
*If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedures==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 liter. A major/large chemical spill requires active assistance from emergency personnel.

==Additional Spill Response Steps:==

'''MINOR CHEMICAL SPILL'''
*Alert people in immediate area of spill.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Open outside windows, if possible.
*Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

'''MAJOR CHEMICAL SPILL'''
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

'''Additional Spill Links:'''
*www.oseh.umich.edu/pdf/chemspil.pdf
*http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling carcinogens.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 13:32, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:18, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:48, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:51, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:48, 13 June 2017 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - B-mercaptoethanol

2019-09-13T19:17:54Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety]]
=Description=
This standard operating procedure (SOP) outlines the handling and use of B-mercaptoethanol (BME). Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling BME.

BME is used as an enzyme reactivator in inhibited systems, a reducing agent in the fluorescent reaction of o-phthaldialdehyde, and amino-acids in alkaline media and is also used to dissociate proteins.

=Potential Hazards=
• BME has a very low odor threshold (0.12-0.64 ppm) and smells similar to the odorant used in natural gas. If the odor becomes widespread, people in nearby areas may suspect a natural gas leak. This can lead to calls to the fire department and/or evacuation of the building, which can be inconvenient and disruptive.

• BME can be toxic if ingested and fatal if inhaled or absorbed through the skin.

• Vapors can irritate the eyes, mucous membranes, and respiratory tract. Symptoms of inhalation exposure may include coughing, sore throat, and/or shortness of breath.

• When BME is heated to decomposition, toxic fumes including sulfur oxides and carbon oxides will be emitted.

• BME is combustible as a liquid or vapor.

• Reactions of BME with strong acids or alkali metals will release flammable hydrogen gas.

=Engineering Controls=
ALWAYS work with BME inside a chemical fume hood or 100% exhausted biological safety cabinet (Class II, Type B2).

=Work Practice Controls=
• BME is incompatible with metals, oxidizing agents, acids, alkalis, calcium hypochlorite, aliphatic amines, and isocyanates.

• Purchase and use in the smallest practical quantities for the experiment being performed.

• Know the location of the nearest fire extinguisher before beginning work.

• Eliminate ignition sources such as open flames and hot surfaces.

• Keep containers closed as much as possible when not in use.

• Be aware of skin absorption as a possible route of exposure. Plan work so that minimal glove contact is expected, and purchase appropriate gloves for cleaning up small spills.

• If glove contact occurs, change gloves immediately.

=Personal Protective Equipment (PPE)=
At a minimum, double-glove using nitrile laboratory gloves and wear a lab coat and safety glasses when working with BME. If there is a possibility of splashing, wear chemical splash goggles and/or a face shield.

=Transportation and Storage=
• Do not store near sources of ignition, oxidizing agents, acids, alkaline compounds, or any other incompatible materials.

• Store BME in a sealed secondary container in a well-ventilated area.

• The container must be tightly closed, resealed, and stored upright to avoid leakage.

• Avoid storing on the floor.

• Transport toxic liquids in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.

• Suitable fire control devices (such as fire extinguishers) must be available at locations where flammable or combustible liquids are stored.

=Waste Disposal=
Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Contact OSEH Hazardous Materials Management at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

=Exposures/Unintended Contact=
If the employee is in need of emergency medical attention, call 911 immediately.

==For an actual chemical exposure/injury:==
• In case of skin contact: Flush the skin with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of eye contact: Flush contaminated eye(s) immediately with copious amounts of water for at least 15 minutes, and then seek medical attention (see below).

• In case of inhalation: Assist conscious persons to an area with fresh, uncontaminated air, and then seek medical attention (see below).

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to Work Connections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the Work Connections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===

U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm

After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021

University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm

Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320

UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedure==
• When a spill occurs, personal safety should always come first.

• Alert and clear everyone in the immediate area where the spill occurred.

Employees in the area should be prepared to clean up minor spills confined to the chemical fume hood. Wearing appropriate gloves (e.g., butyl rubber, Silver Shield®, nitrile), splash goggles, lab coat (and impermeable apron, if available), use an inert absorbent material (sorbent pads, vermiculite, dry sand) to clean up the spill. Do not use combustible materials (i.e. saw dust) to absorb spill. Contaminated PPE and clean-up materials must be placed in a sealed container for pick-up by OSEH-HMM (see waste disposal section for more information).

For large spills, or any spills of BME outside of the fume hood, contact the University of Michigan Division of Public Safety and Security (DPSS) by calling 911.
Additional Spill Links:

• http://oseh.umich.edu/hazardous-waste/spill-response/

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

=Training of Personnel=
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling BME.

=Certification=
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure.

*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:42, 10 November 2016 (UTC)
*[[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:43, 12 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]])
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 21:43, 12 September 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☐ NO
Laboratory Director Revision Date
�
Major Revisions (Tracking purposes only – Do not print as part of SOP)

SOP - Autoclave

2019-09-13T19:17:12Z

Kkistler: KK signed

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the use of autoclaves
Autoclaves are used to sterilize equipment through a process that incorporates high pressure and high temperature.

==Potential Hazards==
Autoclaves pose several potential hazards to labs who frequently use them. Among the physical hazards are heat, steam, and high pressure. Biological hazards, including improperly autoclaved infectious materials, are also present.

Controls for different brands of autoclaves may have their own unique characteristics for loading, load sizes, cycle types and cycle settings. The type of materials to be sterilized will determine the sterilization cycle that will be used. For this reason it is important to read and understand the user’s manual for the specific model of autoclave that is being used prior to first time use. Always ensure that the user’s manual is readily available in case questions or concerns arise during the operation of the autoclave.

==Engineering Controls==
Autoclave monitoring and maintenance is an important aspect of a safe and properly functioning autoclave. Follow the manufacturer’s recommendations for preventive maintenance and ensure that all contractors hired to perform regular maintenance and repairs are approved by the manufacturer. Autoclave operators shall ensure that each autoclave is monitored as follows:

Heat Sensitive Tape Monitoring – Operators shall use heat sensitive sterilization indicator tape for each load to indicate that the load has undergone an effective steam sterilization process.
* Change on autoclave tape means the proper temperature was achieved, but does not mean that temperature was maintained for the proper length of time.
* Place a small piece of autoclave tape on each bottle and tip box.
Biological Indicators – Operators who autoclave medical/biohazardous waste need to do the following:
* At least once a month use a biological indicator such as Bacillus Stearothermophilus (e.g. Prospore2) placed at the center of a load processed under standard operating conditions to confirm the attainment of adequate sterilization conditions.
==Work Practice Controls==
Do not autoclave items containing corrosives, solvents, volatiles or radioactive materials.

===Prior to loading===
* Before using the autoclave, be sure previous users have cleared the autoclave, and that no broken items have been left inside.
* Check to be sure the drain is clear.
* Verify that the interior of the autoclave is in usable condition.

===Loading===
* Carefully load items to be autoclaved, DO NOT overload the autoclave.
* Bottles containing liquids should be placed inside a heat resistant plastic tray (secondary container) containing an inch of water.
* Bottles should not be filled more than 2/3.
* Keep 1-2 inches of space between bottles.
* DO NOT tighten caps on bottles fully.
* Individual glassware pieces should be within a secondary container on a shelf or rack and never placed directly on the autoclave chamber bottom or floor.
* Make sure that the door of the autoclave is fully closed and latched and ensure that the correct cycle for the items being autoclaved has been selected before starting the cycle.

===Opening===
* Wear the proper PPE, including autoclave gloves and close toed shoes when opening the autoclave door after a cycle.
* When the cycle is complete, open the door slowly to minimize exposure to steam. Keep appendages away from the opening.

===When autoclaving liquids:===
* When running an autoclave cycle with liquids, choose the liquid cycle setting.
* To prevent bottles from shattering during the pressurization, the caps of containers with liquids must be loosened before loading.
* Use only borosilicate glass (Pyrex™ or Kimax™).
* Use a secondary container with a solid bottom and walls to contain the contents and catch spills.
* Set a timer to come and unload the autoclave, to ensure it is available for use by other labs.
* Wait 10 minutes after the cycle ends before removing autoclaved liquid load items.
* Let the liquids stand for at least a full hour before touching with ungloved hands. Be sure to let others in the area know that a heat hazard is present.

===When autoclaving dry loads:===
* Add ¼ to ½ inch of water to the tray so that the bottles will heat evenly. Do not fully tighten bottle caps, this could lead to the bottle shattering. Loose caps allow in inside of the bottle to be sterilized.
* Check plastic materials to ensure that they are compatible with being autoclaved.
* Choose the dry porous materials cycle.
* Set a timer to come and unload the autoclave, to ensure it is available for use by other labs.
* Before removing autoclaved items, wait 5 minutes after the cycle ends for loads containing only dry glassware.
* For dry loads, let the glassware cool for a minimum of 15 minutes before touching it with ungloved hands.

==Protective Equipment (PPE)==
Wear the proper PPE, including heat resistant gloves, and close toed shoes when opening the autoclave door after a cycle.
==Waste Disposal==
Place waste in a university approved autoclave bag; avoid over packing of autoclave bags. Do not seal bags or close bottles and other containers tightly. After sterilization cycle, close and seal bag. The sterilized waste can be transported directly to the dumpster by Building Services or laboratory staff without boxing or labeling. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

Discontinue use immediately if an autoclave is not working properly. Post a sign alerting others not to use the autoclave. Mechanical failures need to be attended to by a trained technician, DO NOT try to fix it yourself. Contact facilities to have it repaired.
If you are burned, you should seek medical treatment immediately. Burns to the face, third- degree burns or burns over large areas of the body should be treated as emergencies. (Call 911). Minor burns should be treated by using first aid procedures, including immersing the burn in cool water immediately, removing clothing from the burn area and keeping the injured area cool for at least 5 minutes. Regardless of the severity, notify your Laboratory Director/Supervisor!

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

====TREATMENT FACILITIES:====
====U-M Occupational Health Services -- Campus Employees====
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
====University Health Services -- University students (non-life threatening conditions)====
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
====UMHS Emergency Department -- after clinic hours or on weekends====
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of personnel==
All personnel are required to complete the Autoclave Safety training online through MyLINC (Course BLS013w). All users shall be trained before operating an autoclave; the Laboratory Director/Supervisor is responsible for ensuring that each person in the lab is appropriately trained. All training must be documented and the records maintained in the lab with other safety training certificates. The Laboratory Director/Supervisor is encouraged to use this policy as a guide for training new personnel.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:13, 19 October 2016 (UTC)
* [[User:Davebridges|Davebridges]] ([[User talk:Davebridges|talk]]) 14:53, 8 November 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:11, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:43, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:04, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:09, 12 June 2017 (UTC)
*[[User:Nlambrec|Nlambrec]] ([[User talk:Nlambrec|talk]]) 21:27, 17 February 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? X YES ☐ NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Animal Anesthetics

2019-09-13T19:16:17Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of animal anesthetics including: isoflurane, halothane, enflurane and ether. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls and personal protective equipment when handling animal anesthetics.

==Procedure Location==
The use of animal anesthetics must be performed in an area with good ventilation and controls to capture and exhaust waste anesthetic gases.

==Potential Hazards==
Anesthetic gas and vapor that leaks during medical or research procedures are considered waste anesthetic gases (WAGs). University faculty, staff and students should be aware of the potential risks of WAGs and be advised to take appropriate precautions to reduce exposures. Workers acutely exposed to excessive amounts of anesthetic gas can experience symptoms of drowsiness, headache, nausea, poor judgment and loss of coordination. Chronic symptoms of over-exposure can include liver, kidney and reproductive effects. Safety precautions include the use of an approved gas scavenging system, or using the agent inside a certified chemical fume hood.

The use of ether is not recommended because it is flammable and a mutagen. Be certain that there are no ignition sources present when handling ether. There are restrictions concerning the use of ether with animals. Contact OSEH at (734) 647-1143 concerning the use of ether.

==Engineering Controls==
Anesthetics should not be handled on the bench top without special ventilation or a scavenging system. Anesthetic gas filtering cartridges, snorkel exhaust, fume hoods or other scavenging systems must be used. ULAM provides ventilated procedure rooms designed for use of anesthetic gases in many areas. Fume hoods provide the best protection against exposure to anesthetics in the laboratory and are the preferred ventilation control device when handling greater than 100 cc outside of the original container. Always handle large quantities of ethyl ether in a fume hood due to the flammable nature of the material. If your research does not permit the handling of large quantities of ethyl ether in your fume hood, contact OSEH to review the adequacy of all special ventilation.

Liquid anesthetics administered with a vaporizer must be scavenged. When used properly, vaporizers equipped with activated charcoal canisters (e.g. F/Air) are effective in removing halogenated waste gases. The F/Air canister containing activated charcoal will absorb waste anesthetics for about 12 hours. Note: F/Air Canisters only absorb halogenated anesthetics (e.g. isoflurane, halothane). Immediately before using any anesthesia machine, the F/Air canister should be removed and weighed to evaluate the remaining absorption capacity. The weight should be recorded and dated on the side of the canister. Immediately following the use of an anesthesia machine, the number of hours the machine was in use should be recorded next to the dated weight information.

Canisters that exceed 12 hours of use or 50 grams of accumulated weight (whichever comes first) must be removed and placed in a sealed plastic bag and disposed of as a hazardous waste through OSEH Hazardous Materials Management (HMM) at (734) 763-4568. Thoroughly clean the induction chamber immediately after each use to avoid residual anesthetic waste release into the environment (which can continue to be released for up to three hours). Please refer to the OSEH Guideline entitled Anesthetic Gas Use for additional information on the safe use of anesthetic gases.

==Work Practice Controls==
All anesthetic agents must be clearly labeled with the correct chemical name. Handwritten labels are acceptable; chemical formulas and structural formulas are not acceptable.

Always keep the flow rate of anesthetics to the animal as low as possible during the procedure. High flow rates can increase your exposure to the anesthetic. It is also important to move the point of potential gas release as close to the exhaust system as possible to increase capture of the chemical.

Do not permit containers to remain open on the bench top. The odor thresholds for most liquid anesthetics (except for ether) are well above permissible exposure limits. If you smell the anesthetic the control procedures you are using are inadequate and must be re-evaluated.

==Personal Protective Equipment (PPE)==
Eye protection in the form of safety glasses must be worn at all times when handling anesthetic agents. Ordinary (street) prescription glasses do not provide adequate protection

Single use nitrile or latex gloves must be worn when handling anesthetic agents as well as lab coats, closed toed shoes and pants. Additional protective clothing should be worn if the possibility of skin contact is likely.

==Transportation and Storage==
Ethers form potentially explosive peroxides after exposure to air and light. Since these chemicals are packaged in an air atmosphere, peroxides can form even though the containers have not been opened. Write the date received and date opened on all containers of ether. Opened containers of ether should be discarded within 12 months of opening. Even closed containers of ether must be discarded by the expiration date through OSEH-HMM (734) 763-4568.

Halogenated liquid anesthetic agents (i.e. halothane, enflurane, isoflurane) are not flammable but do have limited shelf life. Be certain to date the chemical when it is opened and to check expiration date before use.

Always purchase the smallest quantity required for use. Ether used for anesthetic purposes should be purchased in the smallest quantity available (typically 150 cc, Fisher Scientific E136-150) due to its short (12 month) shelf life after it is opened.

==Waste Disposal==
Anesthetic agents are hazardous wastes. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, waste collection and for any questions regarding proper waste disposal. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

'''If the employee is in need of emergency medical attention, call 911 immediately.'''

Wash hands and arms with soap and water immediately following any skin contact with anesthetic agents. Flush eyes for 15 minutes following eye contact.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for additional accident and injury reporting information.

==Spill Procedure==
Ether is extremely flammable. If ether is spilled immediately assess and deactivate potential ignition sources. Be prepared for a potential fire and ensure your safety and others first.

Anticipate spills by having the appropriate clean up equipment on hand. Spill materials for anesthetic agents are designed to control the liquid portion of the spill and minimize the production of vapors. Never use paper towels on large spills of anesthetic agents because it exacerbates vapor production.

*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 liter. A major/large chemical spill requires active assistance from emergency personnel.

==Additional Spill Response Steps:==

===MINOR CHEMICAL SPILL===
*Alert people in immediate area of spill.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Open outside windows, if possible.
*Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
*Avoid breathing vapors from spill.
*Confine spill to as small an area as possible.
*Do not wash spill down the drain.
*Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. Contact OSEH-HMM (734) 763-4568 for proper disposal.
*Clean spill area with water.

===MAJOR CHEMICAL SPILL===
*Attend to injured or contaminated persons and remove them from exposure.
*Alert people in the laboratory to evacuate.
*If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
*Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
*Close doors to affected area.
*Post warnings to keep people from entering the area.
*Have person available that has knowledge of incident and laboratory to assist emergency personnel.

==Additional Spill Links:==
• www.oseh.umich.edu/pdf/chemspil.pdf
• http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling animal anesthetics.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:44, 13 October 2016 (UTC)

* [[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 19:34, 10 November 2016 (UTC)

*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:09, 5 June 2017 (UTC)

* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:34, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:38, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:48, 13 June 2017 (UTC)
*[[User:Lgunder|Lgunder]] ([[User talk:Lgunder|talk]]) 18:07, 8 January 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])

Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

SOP - Bleach

2019-09-13T19:10:58Z

Kkistler: KK signed

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the handling and use of bleach. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling bleach.
This document is modified from http://www.oseh.umich.edu/docs/SOP/Bleach.docx

Bleach is a general term that typically refers to an aqueous mixture that has a primary active ingredient of either Sodium Hypochlorite or Calcium Hypochlorite and usually at concentrations of 3 - 10%. Bleach is used primarily as an oxidizing, cleaning, or a bleaching agent, as a disinfectant or within drinking water and waste water purification systems.

Synonyms of bleach include sodium hypochlorite, Clorox, liquid bleach, antiformin, chlorox, Carrel-dakin solution, Chloros, Dakin’s solution, hychlorite, Javelle water.

==Useful Bleach Links==
* http://www.atsdr.cdc.gov/toxfaqs/tfacts184.pdf
* http://www.atsdr.cdc.gov/Mhmi/mmg184.pdf
* http://www.asc.co.id/uplimg/File/03%20zMSDS_NaClO_ASC%20R3.pdf
* http://www.cl2.com/documents/Incompatibility%20Chart.pdf

==Potential Hazards==
* Corrosive. May cause severe irritation or damage to eyes or skin. Vapor may irritate eyes and respiratory tract.
* Incompatible with many chemicals/agents. Hazardous gases (including chlorine and chloramines) may be released if bleach is mixed with an incompatible material.
* Exposure to these gases can cause coughing, shortness of breath, irritation to, or burning of the eyes, nose, and throat, chest pain, wheezing, fluid in the lungs, and nausea. Chlorine can also be absorbed through the skin and cause pain, inflammation, swelling, and blistering.
* Strong oxidizer. May initiate or promote combustion in other materials.

==Occupational Exposure Limits (OELs)==
* MIOSHA: 0.5 ppm Chlorine (from sodium hypochlorite), 8-hour PEL
* MIOSHA: 1 ppm Chlorine (from sodium hypochlorite), 15-minute STEL
* AIHA (WEEL): 2 mg/m3 Sodium Hypochlorite, 15-minute STEL

==Engineering Controls==
* An eyewash/drench hose combination unit must be available in the immediate work area for any work with corrosive materials, including bleach.
* If large quantities will be used, a safety shower will also be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a safety shower if there is not one available.
* Depending on the material’s pH or based on its ability to cause severe tissue damage, e.g., formaldehyde, methylene chloride, methyl ethyl ketone peroxide (MEKP), phenol, etc., the location of the emergency shower and/or emergency eyewash shall be within 25 to 100 feet from the hazardous operation.
* A system of local and/or general exhaust is recommended to keep employee exposures below MIOSHA Permissible Exposure Limits (PEL). Local exhaust ventilation (LEV) is generally preferred because it can control the emissions of the contaminant at its source, preventing dispersion of it into the general work area. Contact OSEH at (734) 647-1143 for a determination of the need for an LEV system, if there is not one available.

==Work Practice Controls==
* ALWAYS review the SDS of both bleach and any chemicals/agents before mixing them with bleach to ensure compatibility. A general list of incompatible materials can be found here.
* NEVER mix bleach with an unknown liquid or unknown residue.
* Do not mix bleach with any compound that is incompatible with oxidizers.
* Purchase bleach in the smallest containers that are practical for lab use.
* Work with the smallest practicable amount and lowest practicable concentration.

==Personal Protective Equipment (PPE)==
In order to select the appropriate PPE for the workplace, a Hazard Assessment is conducted. The hazard assessment determines the hazards and potential hazards associated with a task, machinery, or process. The appropriate PPE for the situation may be subsequently determined. The Hazard Assessment Form may be found here. It may be completed either by the workplace supervisor or the department’s OSEH representative.
* Personal protective equipment is especially important. Wear a buttoned lab coat, safety glasses (that meet the requirements of ANSI/ISEA Z87.1) or goggles if splashing may occur, and gloves for any work with bleach. Depending on the quantities and concentrations used, a face shield, impenetrable, chemical apron and sleeves (or coverall), and special gloves may be recommended.
* Should adequate dilution ventilation or LEV be present, respiratory protection should not be necessary. However, in the absence of these and when MIOSHA PEL, or other published occupational exposures limits (OEL) are anticipated to be exceeded, respiratory protection may be necessary. Contact OSEH at (734) 647-1143 for a determination of the need for a respirator.

==Transportation and Storage==
* Transport concentrated bleach solutions in secondary containment, preferably a polyethylene or other non-reactive acid/solvent bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from metal (unless the metal has a corrosion-proof coating), and do not store under the sink.
* Store away from incompatibles and flammable materials. Always review the SDS of other chemicals in the storage area for compatibility with bleach.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
If the bleach solution is at a household concentration or less, i.e., a maximum 10% concentration of sodium hypochlorite, and was used for disinfecting, limit discharges down a laboratory sink to less than 1-gallon (approx. 3.8 L). If the concentration was >10% and was used for disinfecting, limit the sink disposal to less than 2-cups (approx. 500 mL) of bleach solution. Also flush the drain with adequate volumes of water.

This does not apply to “waste” bleach, e.g., unused or expired bleach. Any leftover, unused product or expired bleach not used as intended by the manufacturer, or used as a disinfectant, needs be collected by OSEH Hazardous Materials Management (HMM) for proper disposal. Contact OSEH-HMM at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to the OSEH Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==
If the employee is in need of emergency medical attention, call 911 immediately.

In general, flush affected eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Eyes: In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Get medical aid.
* Skin: In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get medical aid. Wash clothing before reuse.
* Ingestion: Get medical aid immediately. Do not induce vomiting unless directed to do so by medical personnel. Never give anything by mouth to an unconscious person. If vomiting occurs naturally, have victim lean forward. Wash mouth with water, and then give plenty of milk or water to drink and obtain urgent medical attention.
* Inhalation: If inhaled, remove to fresh air. If not breathing, give artificial respiration. Get medical aid.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities==
* U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
* University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
* UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

==Spill Procedures==
* When a spill occurs, personal safety should always come first.
* Alert and clear everyone in the immediate area where the spill occurred.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 Gallon or 3.5 Liters. A major/large chemical spill requires active assistance from emergency personnel.

For minor (small) spills
* Use proper personal protective equipment as indicated above.
* Absorb the spilled bleach using an absorbent, non-combustible material such as appropriate sorbent pads, sand, or vermiculite. Do not use combustible materials such as sawdust.
* Collect residue, place in container and contact OSEH-HMM (734) 763-4568 for proper disposal.

For major (large) spills
Report large chemical spills greater than 1 gallon or 3.5 liters in corridors or common areas, e.g., hallways, elevators, eating areas, rest rooms, offices, etc., to University of Michigan Division of Public Safety and Security (DPSS) by calling 911.

Additional Spill Response Steps:

===Minor Chemical Spill===
* Alert people in immediate area of spill.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to neutralize corrosives and/or absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===Major Chemical Spill===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

===Additional Spill Links===
* http://www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling bleach.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 16:42, 12 October 2016 (UTC)
* [[User:Ccousz|Ccousz]] ([[User talk:Ccousz|talk]])
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:17, 10 November 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 18:47, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:45, 12 June 2017 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 14:52, 13 June 2017 (UTC)
*[[User:Nlambrec|Nlambrec]] ([[User talk:Nlambrec|talk]]) 21:08, 17 February 2019 (UTC)
*[[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]])
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-07-26

SOP - Irritants

2019-01-07T21:09:26Z

Kkistler: I have read this

[[Category: SOP]]
[[Category: Lab Safety]]

==Description==
This standard operating procedure outlines the handling and use of irritant chemicals. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when handling irritant chemicals.

A chemical irritant is one that is not corrosive but that causes a reversible inflammatory effect on living tissue by chemical action at the site of contact.

==Potential Hazards==
Irritants are chemicals that cause reversible inflammatory effects on living tissue by chemical action at the site of contact. A wide variety of organic and inorganic compounds are irritants; thus, skin contact with all laboratory chemicals should be avoided.
* Substances that may cause eye irritation include: YPD broth, Triton X-100, Trizma base, sodium pyrophosphate decahydrate, sodium fluoride, sodium dodecyl sulfate, poncaeu S, lithium chloride, Luria broth, glycerol, water-soluble dexamethasone, corn oil, and chloroform.
* Substances that may cause skin irritation include: Calcium chloride dehydrate, chloroform, corn oil, water-soluble dexamethasone, luria broth, lithium chloride, Meyer’s hematoxylin, limonene mount, ponceau S, sodium dodceyl sulfate, sodium pyrophosphate decahydrate, trizma base, and YPD broth.
* Substances that may cause respiratory irritation include: YPD broth, trizma base, sodium pyrophosphate decahydrate, sodium dodceyl sulfate, sodium deoxycholate, ponceau S, lithium chloride, luria broth, water soluble dexamethasone, corn oil, and calcium chloride dihydrate.

==Engineering Controls==
Use a properly functioning lab fume hood when handling irritants that can be inhaled (via mist/fume/gas/vapor). If the process does not permit the handing of such materials in a fume hood, contact Occupational Safety and Environmental Health (OSEH) at (734) 647-1143 to review the adequacy of ventilation measures.

==Work Practice Controls==
Handling processes should be designed to minimize the potential for splash, splatter, or other likely scenarios for accidental contact.

Ensure secondary containment and segregation of incompatible chemicals per guidance within the substance-specific storage guidance provided in SDS documentation.
To prevent contact with irritants, be sure to handle them carefully with the proper PPE.

For irritants that are also considered particularly hazardous substances, a designated area shall be established per other applicable SOPs.

==Personal Protective Equipment==
At minimum, safety glasses, lab coat, long pants, and closed toe shoes are to be worn when entering laboratories having hazardous chemicals.
Additionally:
* When handling hazardous chemicals or contacting potentially contaminated surfaces, protective gloves are to be worn. For proper selection of glove material, review the chemical-specific SDS.
* Goggles (not safety glasses) are appropriate for processes where splash or spray is foreseeable.
* For hazardous chemicals that are toxic via skin contact/ absorption, additional protective clothing (i.e., face shield, apron, oversleeves) is appropriate where chemical contact with body/skin is foreseeable.

==Transportation and Storage==
* Transport irritant chemicals in secondary containment, preferably a polyethylene or other non-reactive bottle carrier.
* Store in well-ventilated areas with secondary containment, such as a non-reactive plastic bin.
* Store below eye level.
* Store away from incompatibles. Review the chemical’s SDS for incompatibility information. Also refer to OSEH’s Chemical Storage webpage for more information.
* Avoid storing on the floor. If storing on the floor is necessary, use secondary containment.

==Waste Disposal==
Irritant substances intended for disposal may likely be considered hazardous wastes. Wherever possible, attempt to design research in a manner that reduces the quantity of waste generated. Contact OSEH’s Hazardous Materials Management (HMM) at (734) 763-4568 for waste containers, labels, manifests, and waste collection. Also refer to OSEH’s Hazardous Waste webpage for more information.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

===For an actual chemical exposure/injury,===
* Flush exposed eyes or skin with water for at least 15 minutes, then seek medical attention (see below).
* Consult SDS for guidance on appropriate first aid. Where medical attention is required, be sure to bring along SDS(s) of chemical(s) to aid medical staff in proper diagnosis and treatment.
Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

===TREATMENT FACILITIES:===
===U-M Occupational Health Services -- Campus Employees===
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
===University Health Services -- University students (non-life threatening conditions)===
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
===UMHS Emergency Department -- after clinic hours or on weekends===
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666
Click here for more information.

==Spill Procedure==
Prompt response to chemical spills is critical to protect worker health & safety and to mitigate adverse effects to the environment. For further guidance, refer to chemical-specific MSDS.

A minor (small) chemical spill is one that the laboratory staff is capable of handling safely without the assistance of safety and emergency personnel, i.e., less than 1 gallon or 3.5 liters. A major/large chemical spill requires active assistance from emergency personnel.

===MINOR CHEMICAL SPILL===
* Alert people in immediate area of spill.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Open outside windows, if possible.
* Wear protective equipment, including safety goggles, gloves and long-sleeve lab coat.
* Avoid breathing vapors from spill.
* Confine spill to as small an area as possible.
* Do not wash spill down the drain.
* Use appropriate spill kits/sorbents to absorb spill. Collect contaminated materials and residues and place in container. For powdered chemicals sweep carefully to avoid generation of dust or, if appropriate, use moist sorbent pads or wet the powder with a suitable solvent and then wipe with a dry cloth. Contact OSEH-HMM (734) 763-4568 for proper disposal.
* Clean spill area with water.

===MAJOR CHEMICAL SPILL===
* Attend to injured or contaminated persons and remove them from exposure.
* Alert people in the laboratory to evacuate.
* If spilled material is flammable, turn off ignition and heat sources. Don’t light Bunsen burners or turn on other switches.
* Call University of Michigan Division of Public Safety and Security (DPSS) at 911 immediately for assistance.
* Close doors to affected area.
* Post warnings to keep people from entering the area.
* Have person available that has knowledge of incident and laboratory to assist emergency personnel.

Additional Spill Links:
* www.oseh.umich.edu/pdf/chemspil.pdf
* http://www.oseh.umich.edu/emer-chemical.shtml.

Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.

==Training of Personnel==
All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when handling irritant chemicals.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]]) 21:09, 7 January 2019 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:27, 10 November 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:14, 31 October 2016 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:52, 7 June 2017 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 14:05, 9 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 22:15, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES X NO
Laboratory Director - Dave Bridges Revision Date - 2016-10-19

SOP - Electrophoresis

2019-01-07T21:08:57Z

Kkistler: I have read this

[[ Category: SOP ]]
[[ Category: Lab Safety ]]

==Description==
This standard operating procedure outlines the process of electrophoresis, including the use of equipment and chemicals necessary for the process. Review this document and supply the information required in order to make it specific to your laboratory. In accordance with this document, laboratories should use appropriate controls, personal protective equipment, and disposal techniques when performing electrophoresis.

Electrophoresis uses electrical energy to separate molecules based on their size, structure, and electrical charge.

==Potential Hazards==
Electrophoresis equipment can pose significant electrical hazard in the laboratory. Typical electrophoresis units operating at 100 volts can provide a lethal shock of 25 milliamps. Take the following precautions:

'''Power Supplies:'''
*Inspect to ensure all switches and indicators are in proper working condition and that power cords and leads are undamaged and properly insulated.
*Label equipment with warning: “Danger Electrical Hazard.”
*Connect to ground fault circuit interrupters (GFCIs).
*Use 3-prong plugs.
*Use power supplies with safety features that detect no-load, overload, sudden load change, short circuit, arc or ground leak, etc.

'''Connecting Leads:'''
*Turn off main power supply before connecting or disconnecting electrical leads.
*With dry gloved hands, connect one lead at a time using one hand only.
*Be sure that leads/banana plugs are fully seated.

'''Using Equipment:'''
*Don’t run equipment unattended.
*Keep equipment clear of unintentional grounding points and conductors (e.g., sinks or other water sources, metal plates, jewelry, aluminum foil, pipes or other electrical/metal equipment).
*Gel chamber must have a lid or cover with safety interlocks to prevent accidental contact with energized electrodes or buffer solutions.
*Gel chamber exterior must be dry with no spilled solutions. Check for leaks.
*Switch off all power and unplug the leads before opening the gel chamber lid or reaching inside the gel chamber.
�
'''Hazardous chemicals'''
Commonly used in conjunction with electrophoresis work include:
*Ethidium bromide – mutagen, irritant
*Acrylamide – carcinogen, neurotoxin, irritant
*Phenol – corrosive, toxic
*Chloroform – suspect carcinogen, toxic

Always review the Safety Data Sheet prior to working with any hazardous material.
Laboratory personnel may be exposed to thermal hazards posed by liquefied gels.

'''Ultraviolet''' (UV) light boxes are often used in visualizing ethidium bromide gels and pose potential exposures to UV radiation.

==Engineering and Work Practice Controls==
*Read and follow manufacturer’s instructions for electrophoresis equipment.
*Prepare Standard Operating Procedure relevant to health and safety.
*Consult with Lab Director prior to initial use of electrophoresis equipment. Discussion should include special hazards and safety precautions.
*Measure, mix and handle all hazardous powdered chemicals or gel prep mixtures with hazardous components (e.g., acrylamide monomer, ethidium bromide, phenol, ammonium persulfate, and formaldehyde) in a fume hood.
*Purchase pre-made gels or pre-mixed acrylamide and ethidium bromide solutions instead of making your own.
*Consider using ethidium bromide substitutes.
*Exercise caution when using microwave to liquefy gels – don’t use sealed containers, beware of superheated liquids that may froth up unexpectedly. Let hot gel preps cool to 50°-60°C before adding ethidium bromide or pouring into trays. Wear insulated gloves and point the flask opening away from you.

==Protective Equipment==
*Wear lab coat with fully extended sleeves, safety glasses or splash goggles if appropriate, nitrile gloves (latex is not effective), pants, and closed-toe shoes.
*Wear appropriate skin and eye protection for UV radiation work.

==Waste Disposal==
'''Hazardous Waste Management:''' Handle and store hazardous waste following the guidelines above for work practice controls, transportation and storage. Contact OSEH Hazardous Materials Management (HMM) at 734-763-4568 with questions and to schedule a pickup of hazardous waste. For more information regarding chemical waste, refer to the HMM section on OSEH’s website at http://www.oseh.umich.edu/hazmats/chemical.shtml.

'''Non-Hazardous Waste Management:''' Some gels may be considered non-hazardous but still present a health hazard if untreated or discarded into the normal trash. Contact HMM for proper collection and disposal.

==Exposures/Unintended Contact==

If the employee is in need of emergency medical attention, call 911 immediately.

For an actual chemical exposure/injury:
*Remove contaminated clothing. Flush exposed eyes or skin with water for at least 15 minutes. Seek medical attention (see below).
*For inhalation exposure, remove all persons from the contaminated area. Get medical aid.
*If an ambulance is needed, call the University of Michigan Division of Public Safety and Security (DPSS) at 911 to request assistance.

Contact OSEH for advice on symptoms of chemical exposure, or assistance in performing an exposure assessment.

Report all work related accidents, injuries, illnesses or exposures to WorkConnections within 24 hours by completing and submitting the Illness and Injury Report Form. Follow the directions on the WorkConnections website Forms Instructions to obtain proper medical treatment and follow-up.

Complete the OSEH Laboratory Incident and Near-Miss Report form.

==Treatment Facilities:==
*U-M Occupational Health Services -- Campus Employees
Mon-Fri 7:30 am - 4:30 pm
After hours - go to UM Hospital Emergency Dept. – Urgent Care Clinic
C380 Med Inn building
1500 East Medical Center Drive, Ann Arbor (734) 764-8021
*University Health Services -- University students (non-life threatening conditions)
Mon-Fri 8 am – 4:30 pm, Sat 9 am – 12 pm
Contact for current hours as they may vary
207 Fletcher Street, Ann Arbor (734) 764-8320
*UMHS Emergency Department -- after clinic hours or on weekends
1500 East Medical Center Drive, Ann Arbor, (734) 936-6666

Click here for more information.

==Spill Procedure==
*When a spill occurs, personal safety should always come first.
*Alert and clear everyone in the immediate area where the spill occurred.
*Follow spill procedures described in the SOP and SDS for the particular chemical spilled.

'''Report all emergencies, suspicious activity, injuries, spills, and fires to the University of Michigan Division of Public Safety and Security (DPSS) by calling 911 or texting 377911. Register with the University of Michigan Emergency Alert System via Wolverine Access.
'''

==Training of Personnel==

All personnel are required to complete the General Laboratory Safety Training session (BLS025w or equivalent) via OSEH’s My LINC website. Furthermore, all personnel shall read and fully adhere to this SOP when performing electrophoresis.

==Certification==
I have read and understand the above SOP. I agree to contact my Supervisor or Lab manager if I plan to modify this procedure. Sign by logging in and typing <nowiki>* ~~~~</nowiki> in the list below:

* [[User:Kkistler|Kkistler]] ([[User talk:Kkistler|talk]]) 21:08, 7 January 2019 (UTC)
*[[User:Reddj|Reddj]] ([[User talk:Reddj|talk]]) 15:00, 13 October 2016 (UTC)
*[[User:Iharvey|Iharvey]] ([[User talk:Iharvey|talk]]) 21:19, 10 November 2016 (UTC)
*[[User:Mollyec|Mollyec]] ([[User talk:Mollyec|talk]]) 16:15, 31 October 2016 (UTC)
*[[User:Pfeiferl|Pfeiferl]] ([[User talk:Pfeiferl|talk]]) 18:10, 5 June 2017 (UTC)
* [[User:Snyderds|Snyderds]] ([[User talk:Snyderds|talk]]) 19:12, 7 June 2017 (UTC)
* [[User:Elhabbal|Elhabbal]] ([[User talk:Elhabbal|talk]]) 21:57, 12 June 2017 (UTC)
Prior Approval required – Is this procedure hazardous enough to warrant prior approval from the Laboratory Director? ☐ YES ☒ NO

Laboratory Director - Dave Bridges Revision Date - 2016-10-13

Western Blotting

2019-01-07T20:20:43Z

Kkistler: Corrections to protocol

==SOP==
*Irritant, Electrophoresis

==Materials==
*Transfer Buffer (200 mL Methanol, 100 mL 10X [[ Transfer Buffer ]] to final 1L volume)
*Transfer Apparatus, either Bio-Rad or Invitrogen

==Protocol==
#Turn on heat block to 85 degrees
#Run SDS-PAGE gel using [[ SDS-PAGE Running Buffer ]] and prepare diluted transfer buffer
## Use a prepared 4-12% tris gel (in the 4 degree). Remove it from the packaging, remove the white strip of tape from the bottom back, and gently pull the comb out and rinse with water.
##Load into gel tank. Fill with SDS page Running buffer (1X) to the fill line in the front and halfway up the back
##Boil sample at 85 degrees for ~3 min
##Load 3 microliters of protein ladder (purple) (in the 4 degree), and 10 microliters of each sample into separate wells.
## Place top on tank, plug into power source and run at 125 Volts until samples and ladder reach the bottom of the gel
#Make sandwich (black side, sponge, filter paper, gel, nitrocellulose, filter paper, sponge, clear side), ensuring no bubbles between layers with black piece on bottom and layer as above. Place in apparatus so that the black sandwich touches the black transfer piece. Fill with transfer buffer.
#Transfer 4h at 75V (in cold room) or overnight at 35V (room temp).
#Stain for total protein with Revert total protein stain on rocker for 5 minutes --when finished pour total protein stain back in bottle for later use!
#Rinse twice in revert wash solution (60ml MeOH, 13.4 ml Acetic Acid, 126.6 ml Water)
#Scan using licor for total protein, which will be used to normalize the blot
#Rinse nitrocellulose in revert reversal solution for at least 5 and no more than 10 minutes until nitrocellulose appears clear again (.2g NaOH, 60ml MeOH, 140ml Water)
#Rinse nitrocellulose in 2% BSA (2g BSA in 100ml TBST, stored in fridge) for 1 hour
#Incubate with primary antibody (check for dilution) in 2% BSA for >1h
#Wash blot every 5 minutes for 15 min with TBST.
#Incubate with appropriate secondary antibody (10 000X) for 45min-1h (20 ml 2% BSA 1ul of both secondary antibodies, all found in fridge Ab stocks with blue dots on top)
#Wash blot every 5 minutes for 15 min with TBST.
#Rinse once or twice with double distilled water
#Prepare ECL by mixing one volume of the black bottle solution with one volume of the white bottle solution (be careful not to cross-contaminate ECL bottles with wrong solution).
#Drain excess buffer from blot and cover with ECL for about a minute
#Drain excess ECL from blot, cover with saran wrap and expose film

==If Using LiCor==
#Start -> New -> Scan Image -> Login -> Peloquin -> Password Located in Desk -> Select Dimensions -> Start Scan

[[ Category: Western Blotting ]]

Western Blotting

2019-01-07T20:16:27Z

Kkistler: Added SOP

==SOP==
*Irritant, Electrophoresis

==Materials==
*Transfer Buffer (200 mL Methanol, 100 mL 10X [[ Transfer Buffer ]] to final 1L volume)
*Transfer Apparatus, either Bio-Rad or Invitrogen

==Protocol==
#Run SDS-PAGE gel using [[ SDS-PAGE Running Buffer ]] and prepare diluted transfer buffer
## Use a prepared 5-12% tris gel (in the 4 degree). Remove it from the packaging, remove the white strip of tape from the bottom back, and gently pull the comb out and rinse with water.
##Load into gel tank. Fill with SDS page Running buffer (1X) to the fill line in the front and halfway up the back
##Load 3 microliters of protein ladder (purple), and 10 microliters of each sample into separate wells.
## Place top on tank, plug into power source and run at 125 Volts until samples and ladder reach the bottom of the gel
#Make sandwich (black side, sponge, filter paper, gel, nitrocellulose, filter paper, sponge, clear side), ensuring no bubbles between layers with black piece on bottom and layer as above. Place in apparatus so that the black sandwich touches the black transfer piece. Fill with transfer buffer.
#Transfer 4h at 75V (in cold room) or overnight at 35V (room temp).
#Stain for total protein with Revert total protein stain on rocker for 5 minutes --when finished pour total protein stain back in bottle for later use!
#Rinse twice in revert wash solution (60ml MeOH, 13.4 ml Aceditc Acid, 126.6 ml Water)
#Scan using licor for total protein, which will be used to normalize the blot
#Rinse nitrocellulose in revert reversal solution for at least 5 and no more than 10 minutes until nitrocellulose appears clear again (.2g NaOH, 60ml MeOH, 140ml Water)
#Rinse nitrocellulose in 2% BSA (2g BSA in 100ml TBST, stored in fridge) for 1 hour
#Incubate with primary antibody (check for dilution) in 2% BSA for >1h
#Wash blot every 5 minutes for 15 min with TBST.
#Incubate with appropriate secondary antibody (10 000X) for 45min-1h (20 ml 2% BSA 1ul of both secondary antibodies, all found in fridge Ab stocks with blue dots on top)
#Wash blot every 5 minutes for 15 min with TBST.
#Rinse once or twice with double distilled water
#Prepare ECL by mixing one volume of the black bottle solution with one volume of the white bottle solution (be careful not to cross-contaminate ECL bottles with wrong solution).
#Drain excess buffer from blot and cover with ECL for about a minute
#Drain excess ECL from blot, cover with saran wrap and expose film

==If Using LiCor==
#Start -> New -> Scan Image -> Login -> Peloquin -> Password Located in Desk -> Select Dimensions -> Start Scan

[[ Category: Western Blotting ]]