Standard Operating Procedure
Hydrazine
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box below 2) lab specific protocol/procedure is added to the protocol/procedure section and
3) SOP has been signed and dated by the PI and relevant lab personnel.
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Laboratory Safety Manual and Chemical Hygiene Plan.
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Type of SOP:
☐Process
☒Hazardous Chemical
☒ Hazardous Class
Purpose
Hydrazine is mainly used as a foaming agent in preparing polymer foams, but significant applications also
include its uses as a precursor to polymerization catalysts and pharmaceuticals. Additionally, hydrazine is
used in various rocket fuels and to prepare the gas precursors used in air bags (Sodium azide, the gasforming agent in air bags, is produced from hydrazine by reaction with sodium nitrite). Hydrazine is used
within both nuclear and conventional electrical power plant steam cycles to control concentrations of
dissolved oxygen in an effort to reduce corrosion. Hydrazine is a useful building block in organic synthesis
of pharmaceuticals and pesticides. It is used as an oxygen scavenger and corrosion inhibitor in water
boilers and heating systems. Some color photographic processes also use a weak solution of hydrazine
as a stabilizing wash, as it scavenges dye coupler and unreacted silver halides. In addition, a
semiconductor deposition technique using hydrazine has recently been demonstrated, with possible
application to the manufacture of thin-film transistors used in Liquid Crystal Displays (LCD systems).
Hydrazine is a colorless flammable liquid with an ammonia-like odor. Hydrazine is highly toxic and
dangerously unstable unless handled in solution. It is a select carcinogen and a reproductive toxin. It is
air, moisture, heat, light and friction sensitive chemical. If not stored and handled properly, this can pose a
serious threat to the health and safety of laboratory personnel, emergency responders and chemical
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waste handlers. In laboratories hydrazine is most frequently found bound to biotin and fluorescent
markers which are used to label specific proteins and cells.
At the low concentration of hydrazine found in these markers there are no known health side effects.
However be aware that because these markers are hydrazine derivatives, they may lead to irritation if
inhaled or to irritation when they come in contact with the skin or mucous membranes. These markers are
prepared by various manufacturers and are used for cell and protein labeling.
Physical & Chemical Properties/Definition of Chemical Group
CAS#: 302-01-2
Class: Click here to enter text.
Molecular Formula: N2H4
Form (physical state): Liquid
Color: Colorless
Boiling point: 113°C
Potential Hazards/Toxicity
Danger!
Strong reducing agent
Fire and explosion risk in contact with oxidizing agents
May be fatal if absorbed through the skin
Causes eye and skin burns
Causes digestive and respiratory tract burns
Flammable liquid and vapor
Harmful if inhaled or swallowed
May cause allergic skin reaction
Cancer suspect agent
May cause blood abnormalities
May cause liver and kidney damage
Potential Health Effects
Eye: May cause irreversible eye injury. Exposure to the vapors or liquid may cause temporary blindness.
Causes severe eye irritation and burns.
Skin: May be fatal if absorbed through the skin. Prolonged and/or repeated contact may cause irritation
and/or dermatitis. May cause skin sensitization, an allergic reaction, which becomes evident upon reexposure to this material. Contact with liquid is corrosive and causes severe burns and ulceration.
Contact with the skin may dissolve hair.
Ingestion: Harmful if swallowed. Causes gastrointestinal irritation with nausea, vomiting and diarrhea.
May cause liver and kidney damage. Causes digestive tract burns with immediate pain, swelling of the
throat, convulsions, and possible coma. Exposure may cause anemia and other blood abnormalities.
Inhalation: Harmful if inhaled. Irritation may lead to chemical pneumonitis and pulmonary edema. May
cause liver and kidney damage. Causes severe irritation of upper respiratory tract with coughing, burns,
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breathing difficulty, and possible coma. Vapors may cause dizziness, nausea, itching, burning, and
swelling of the eyes.
Chronic: Repeated inhalation may cause chronic bronchitis. May cause cancer according to animal
studies. Repeated exposure may cause sensitization dermatitis. May cause reproductive and fetal
defects.
Permissible Exposure Limit (PEL) - OSHA Vacated PELs: 0.1 ppm TWA; 0.1 mg/m3 TWA
National Fire Protection Association – NFPA ratings:
Health: 3
Flammability: 3
Reactivity: 3
Personal Protective Equipment (PPE)
Respirator Protection
Respirators should be used only under any of the following circumstances:
 As a last line of defense (i.e., after engineering and administrative controls have been
exhausted).
 When Permissible Exposure Limit (PEL) has exceeded or when there is a possibility that PEL will
be exceeded.
 Regulations require the use of a respirator.
 An employer requires the use of a respirator.
 There is potential for harmful exposure due to an atmospheric contaminant (in the absence of
PEL)
 As PPE in the event of a chemical spill clean-up process
Lab personnel intending to use/wear a respirator mask must be trained and fit-tested by EH&S. This is a
regulatory requirement. (https://www.ehs.ucla.edu/ep/ih/resp)
Hand Protection
Handle with Neoprene/Nitrile gloves.
NOTE: Consult with your preferred glove manufacturer to ensure that the gloves you plan on using are
compatible with Hydrazine.
Refer to glove selection chart from the links below:
http://www.ansellpro.com/download/Ansell_8thEditionChemicalResistanceGuide.pdf
OR
http://www.allsafetyproducts.com/glove-selection-chart-chemical-breakthrough-ratings.html
OR
http://www.showabestglove.com/site/default.aspx
OR
http://www.mapaglove.com/
Gloves must be inspected prior to use.
Use proper glove removal technique (without touching glove's outer surface) to avoid skin contact with
hydrazine.
Wash with warm water and soap, dry hands.
Eye Protection
Safety goggles.
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Skin and Body Protection
Lab coats should be worn. These laboratory coats must be appropriately sized for the individual and be
buttoned to their full length. Laboratory coat sleeves must be of a sufficient length to prevent skin
exposure while wearing gloves. Full length pants and close-toed shoes must be worn at all times by all
individuals that are occupying the laboratory area. The area of skin between the shoe and ankle should
not be exposed.
Hygiene Measures
Avoid contact with skin, eyes and clothing. Wash hands before breaks and immediately after handling
hydrazine.
Engineering Controls
Store & handle inside a glove box (under an inert atmosphere – preferably under Nitrogen). For additional
information, please refer to the MSDS (i.e., sections on ‘Stability & Reactivity’ and ‘Handling & Storage’)
from the same manufacturer.
First Aid Procedures
If inhaled
Get medical aid immediately. Remove from exposure and move to fresh air immediately. If breathing is
difficult, give oxygen. Do NOT use mouth-to-mouth resuscitation. If breathing has ceased apply artificial
respiration using oxygen and a suitable mechanical device such as a bag and a mask.
In case of skin contact
Get medical aid immediately. Immediately flush skin with plenty of water for at least 15 minutes while
removing contaminated clothing and shoes. Discard contaminated clothing in a manner which limits
further exposure.
In case of eye contact
Immediately flush eyes with plenty of water for at least 15 minutes, occasionally lifting the upper and
lower eyelids. Get medical aid immediately. Do NOT allow victim to rub eyes or keep eyes closed.
If swallowed
Do not induce vomiting. If victim is conscious and alert, give 2-4 cupfuls of milk or water. Never give
anything by mouth to an unconscious person. Get medical aid immediately.
Special Handling and Storage Requirements
Handling
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Wash thoroughly after handling.
Remove contaminated clothing and wash before reuse.
Ground and bond containers when transferring material (i.e., if in a metal container). For safety
reasons, transfer from glass to glass container using proper techniques (often purged with
nitrogen).
Never add water to this product.
Avoid shock and friction.
Use spark-proof tools and explosion proof equipment.
Do not get in eyes, on skin, or on clothing.
Empty containers retain product residue, (liquid and/or vapor), and can be dangerous.
Keep container tightly closed.
Do not ingest or inhale.
Do not pressurize, cut, weld, braze, solder, drill, grind, or expose empty containers to heat,
sparks or open flames.
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Keep away from heat, sparks and flame.
Use only with adequate ventilation or respiratory protection.
Storage
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Keep locked up. Keep container dry.
Keep away from heat, sparks & flame.
Keep away from sources of ignition.
Keep away from acids and moisture.
Do not store in direct sunlight or strong incandescent light.
Storage under a nitrogen blanket has been recommended OR inside a glove box (under inert
atmosphere).
Isolate from oxidizing materials and acids.
NOTE
 All hydrazine containers must be stored using secondary container (tray/tub) with proper
signage/caution label.
 Hazard communication: Warning/Caution label is required on each hydrazine container,
secondary containment and designated storage cabinets/areas. Warning/Caution label must state
the following:
“CARCINOGEN / CANCER HAZARD or SUSPECT CANCER AGENT & Reproductive Toxin”
Chemical Stability: Thermally unstable.
Conditions to Avoid: Light, ignition sources, moisture, temperatures above 150°C.
Incompatibility
 Substance is highly reactive reducing agent.
 Incompatible with oxidizing agents (including air), acids, and some metal oxides and metals.
Substance may spontaneously ignite in air when in contact with porous materials.
 Ignites on contact with dinitrogen oxide and tetroxide, hydrogen peroxide, tetryl, and nitric acid.
Explodes on contact with dicyanofurazan, n-halomides, potassium, silver compounds, sodium
hydroxide, titanium compounds, and trioxygen difluoride.
 Explosive compounds may result from contact with air - chloromethylnitrobenzene, lithium
perchlorate, metal salts, methanol + nitromethane, sodium, and sodium perchlorate.
 Also incompatible with barium oxide or calcium oxide, benzeneseleninic acid or anhydride,
calcium, carbon dioxide + stainless steel, 1-chloro-2,4-dinitrobenzene, cotton waste + heavy
metals, (difluoroamino) difluoroacetonitrile, iodine pentoxide, rust, ruthenium(III) oxide,
thiocarbonyl azide thiocyanate,
Hazardous Decomposition Products: Oxides of nitrogen, ammonia and/or derivatives.
Spill and Accident Procedure
Fire Extinguishers and Extinguishing Media
General Information:
Wear a Self-Contained Breathing Apparatus (SCBA) mask and full protective gear.
Vapors can travel to a source of ignition and flash back.
Is a flammable liquid and vapor.
Auto-ignition temperature varies based upon contact surface.
Vapors may be heavier than air.
They can spread along the ground and collect in low or confined areas.
May be ignited by heat, sparks, and flame.
Vapors may form an explosive mixture with air.
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Containers may explode when heated.
Extinguishing Media:
For large fires, use water spray, fog, or alcohol-resistant foam.
Use water spray to cool fire-exposed containers.
Do NOT get water inside containers.
For small fires, use dry chemical, carbon dioxide, or water spray.
Cool containers with flooding quantities of water until well after fire is out.
Flash Point: 38 deg C (100.4 deg F)
Auto-ignition Temperature: 270 deg C (518.00 deg F)
Explosion Limits, Lower: 2.9%
Upper: 98%
Chemical Spill Dial 911 and x59797
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Remove all sources of ignition.
Absorb spill using an absorbent, non-combustible material such as earth, sand, or vermiculite.
Do not use combustible materials such as sawdust.
Do not get water inside containers.
Approach spill from upwind (if outdoors).
Use water spray to cool and disperse vapors, protect personnel, and dilute spills to form
nonflammable mixtures.
Control runoff and isolate discharged material for proper disposal (if outdoors).
Spill – Assess the extent of danger. Help contaminated or injured persons. Evacuate the spill area.
Avoid breathing vapors. If possible, confine the spill to a small area using a spill kit or absorbent material.
Keep others from entering contaminated area (e.g., use caution tape, barriers, etc.).
Chemical Spill on Body or Clothes – Remove clothing and rinse body thoroughly in emergency shower
for at least 15 minutes. Seek medical attention. Notify supervisor and EH&S at x59797 immediately.
Chemical Splash Into Eyes – Immediately rinse eyeball and inner surface of eyelid with water from the
emergency eyewash station for 15 minutes by forcibly holding the eye open. Seek medical attention.
Notify supervisor and EH&S at x59797 immediately.
Medical Emergency Dial 911 or x52111
Life Threatening Emergency, After Hours, Weekends And Holidays – Dial 911 (or 310-825-1491 from
cell phone) or contact the Ronald Reagan UCLA Medical Center (emergency room) directly at x52111
(located at 757 Westwood Plaza, enter from Gayley Avenue). Note: All serious injuries must be reported
to EH&S at x59797 within 8 hours.
Non-Life Threatening Emergency – Go to the Occupational Health Facility (OHF), x56771, CHS room
67-120 (This is on the 6th floor, 7th corridor, room 120. Enter through the School of Dentistry on Tiverton
Drive and proceed to the “O” elevator to the 6th floor.)Hours: M - F, 7:30 a.m. to 4:30 p.m. At all other
times report to Ronald Regan UCLA Medical Center (emergency room) at x52111. Note: All serious
injuries must be reported to EH&S at x59797 within 8 hours.
Needle stick/puncture exposure (as applicable to chemical handling procedure) – Wash the affected
area with antiseptic soap and warm water for 15 minutes. For mucous membrane exposure, flush the
affected area for 15 minutes using an eyewash station. Page the needle stick nurse by dialing 231 from a
campus phone, enter 93333 when prompted and then enter your extension. Hours: M – F, 8:00 a.m. to
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4:00 p.m. At all other times report to Ronald Regan UCLA Medical Center (emergency room) at x52111.
Note: All needle stick/puncture exposures must be reported to EH&S at x59797 within 8 hours.
Decontamination/Waste Disposal Procedure
Hydrazine is listed as Extremely Hazardous (EH) Substance by the State. Even the containers
(irrespective of the size) which once held hydrazine must be disposed of as hazardous waste with an online hazardous waste tag affixed on the container. (http://map.ais.ucla.edu/go/1002774)
All dry hazardous waste must be double bagged (use only transparent bags) and affixed with an on-line
waste tag. Note: Black, Red, Orange or Yellow bags are prohibited by EH&S Hazardous Materials
Division, for hazardous waste disposal.
General hazardous waste disposal guidelines:
Label Waste
 Affix an on-line hazardous waste tag on all waste containers using the Online Tag Program
http://otp.ucop.edu/ as soon as the first drop of waste is added to the container
Store Waste
 Store hazardous waste in closed containers, in secondary containment and in a designated
location
 Double-bag dry waste using transparent bags http://map.ais.ucla.edu/go/1002774
 Waste must be under the control of the person generating & disposing of it
Dispose of Waste
 Dispose of regularly generated chemical waste within 90 days
 Call EH&S at x61887 for questions
 Empty Containers
o Dispose as hazardous waste if it once held extremely hazardous waste (irrespective of
the container size) http://ehs.ucla.edu/Pub/ExtremelyHazardousWaste.pdf
o Consult waste pick-up schedule
http://ehs.ucla.edu/pub/HazWaste%20Pickup%20Schedule.pdf
Prepare for transport to pick-up location
 Check on-line waste tag
 Write date of pick-up on the waste tag
 Use secondary containment
Safety Data Sheet (SDS) Location
Hardcopy and electronic copy must be available for hydrazine.
Online SDS can be accessed at http://msds.ehs.ucla.edu.
Protocol/Procedure
Amount covered
0-50 mg lyophilized powder
0-400 ul reconstituted 50 mg in DMSO stored at -20 C
Temperature range covered
-22°C - 26°C
Labeling Glycoproteins with Biotin Hydrazide Reagents
Materials Required
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Biotin Hydrazide Solution: 50 mM biotin hydrazide reagent in dimethylsulfoxide (DMSO, Product
No. 20684). Prepare a volume sufficient to achieve the desired final concentration in step B.4.
Excess (unused) dissolved reagent may be stable, but this has not been verified.
Note:Biotin-PEG4-Hydrazide is a hygroscopic solid that is difficult to weigh and dispense. To
facilitate handling, make a stock solution immediately before first use by dissolving the entire
contents of the vial (50 mg) in 396 μl of dry (anhydrous, molecular sieve-treated) organic solvent
such as DMSO. Store the resulting 250 mM stock solution of Biotin-PEG4-Hydrazide at -20°C,
and warm the vial fully to room temperature before opening for use to prevent moisture
condensation, which may decrease its shelf-life.
Oxidation Buffer: 0.1 M sodium acetate buffer, pH 5.5
Sodium meta-periodate solution: 20 mM sodium meta-periodate in Oxidation Buffer. Prepare
solution immediately before use in amber vial or other light-protecting vessel.
Coupling Buffer: 0.1 M sodium phosphate, 0.15 M NaCl, pH 7.2 (PBS, Product No. 28372) or
other neutral or slightly alkaline, non-amine buffer
Glycoprotein Solution: 2 mg/ml of glycoprotein in Oxidation Buffer
Dialysis cassette or desalting
Procedure
1. Add 1 ml of cold sodium meta-periodate solution to 1 ml of cold glycoprotein solution; mix well
and then protect reaction vessel from light and incubate mixture for 30 minutes on ice or at 4°C.
2. Remove excess periodate and exchange the sample buffer by dialysis against coupling buffer or
gel filtration through a desalting column that has been equilibrated with coupling buffer.
3. Add 1 part prepared 50 mM Biotin Hydrazide Solution to 9 parts oxidized and buffer-exchanged
sample (results in 5 mM Biotin Hydrazide); mix for 2 hours at room temperature.
4. Separate the biotinylated molecule from non-reacted material by dialysis or gel filtration (desalting
column). Biotinylated samples may be stored using the same conditions as for the nonbiotinylated sample
NOTE
Any deviation from this SOP requires approval from PI.
Documentation of Training (signature of all users is required)
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Prior to conducting any work with Hydrazine, designated personnel must provide training to his/her
laboratory personnel specific to the hazards involved in working with this substance, work area
decontamination, and emergency procedures.
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The Principal Investigator must provide his/her laboratory personnel with a copy of this SOP and a
copy of the SDS provided by the manufacturer.
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The Principal Investigator must ensure that his/her laboratory personnel have attended appropriate
laboratory safety training or refresher training within the last one year.
Principal Investigator SOP Approval
Print name __________________________Signature___________________________
Approval Date:
I have read and understand the content of this SOP:
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